"``ChimericJJcoronavirus-likeJJparticlesNNScarryingVBGsevereJJacuteJJrespiratoryJJsyndromeNNcoronavirus (NNSCoV)NNSNNproteinNNprotectVBPmiceNNSagainstINchallengeNNwithINSCoV",NN_SPpunctamodamodnsubjaclamodamodamodcompoundcompoundagentcompounddobjdobjcasenmodcasepunct
displaCy
"Chimeric coronavirus-likeGENE_OR_GENE_PRODUCT particles carrying severe acute respiratory syndrome coronavirusORGANISM (SCoV) S protein protect miceORGANISM against challenge with SCoVSIMPLE_CHEMICAL",
displaCy
"``WePRPtestedVBDtheDTefficacyNNofINcoronavirus-likeJJparticles (NNSVLPs)NNSforINprotectingVBGmiceNNSagainstINsevereJJacuteJJrespiratoryJJsyndromeNNcoronavirus (NNSCoV)NNinfection.NNpunctnsubjdetdobjcaseamodnmodapposmarkadvcldobjcaseamodamodamodcompoundcompoundapposnmod
displaCy
"We tested the efficacy of coronavirus-like particlesGENE_OR_GENE_PRODUCT (VLPsORGANISM) for protecting miceORGANISM against severe acute respiratory syndrome coronavirusORGANISM (SCoVSIMPLE_CHEMICAL) infection.
displaCy
CoexpressionNNofINSCoVNNSNNproteinNNandCCE,NNMNNandCCNNNproteinsNNSofINmouseNNhepatitisNNvirusNNinIN293CDTNNorCCCHONNcellsNNSresultedVBDinINtheDTefficientJJproductionNNofINchimericJJVLPsNNScarryingVBGSCoVNNSNNprotein.NNnsubjcaseamodcompoundnmodcccompoundconjccconjconjcasecompoundcompoundnmodcasenummodnmodccconjagentcasedetamodnmodcaseamodnmodaclamodcompounddobj
displaCy
Coexpression of SCoV SGENE_OR_GENE_PRODUCT protein and E, M and N proteins of mouse hepatitis virusORGANISM in 293T or CHO cellsCELL resulted in the efficient production of chimeric VLPsCELL carrying SCoV SGENE_OR_GENE_PRODUCT protein.
displaCy
Balb/cNNmiceNNSinoculatedVBNwithINaDTmixtureNNofINchimericJJVLPsNNSandCCalumVBNtwiceRBatINanDTintervalNNofINfourCDweeksNNSwereVBDprotectedVBNfromINSCoVNNchallenge,NNasINindicatedVBNbyINtheDTabsenceNNofINinfectiousJJvirusNNinINtheDTlungs.NNScompoundnsubjpassaclcasedetnmodcaseamodnmodccconjadvmodcasedetnmodcasenummodnmodauxpasscaseamodnmodmarkadvclcasedetnmodcaseamodnmodcasedetnmod
displaCy
Balb/c miceORGANISM inoculated with a mixture of chimeric VLPsCELL and alumSIMPLE_CHEMICAL twice at an interval of four weeks were protected from SCoVSIMPLE_CHEMICAL challenge, as indicated by the absence of infectious virus in the lungsORGAN.
displaCy
TheDTsameJJgroupsNNSofINmiceNNShadVBDhighJJlevelsNNSofINSCoV-specificJJneutralizingVBGantibodies,NNSwhileINmiceNNSinINtheDTnegativeJJcontrolNNgroups,NNSwhichWDTwereVBDnotRBimmunizedVBNwithINchimericJJVLPs,NNSfailedVBDtoTOmanifestVBneutralizingVBGantibodies,NNSsuggestingVBGthatINSCoV-specificJJneutralizingVBGantibodiesNNSareVBPimportantJJforINtheDTsuppressionNNofINviralJJreplicationNNwithinINtheDTlungs.NNSdetamodnsubjcasenmodamoddobjcaseamodamodnmodmarknsubjcasedetamodcompoundnmodnsubjpassauxpassnegparataxiscaseamodnmodadvclcaseamodamodnmodxcompmarkamodamodnsubjpredetccompcasedetnmodcaseamodnmodcasedetnmod
displaCy
The same groups of miceORGANISM had high levels of SCoV-specific neutralizing antibodies, while miceORGANISM in the negative control groups, which were not immunized with chimeric VLPs, failed to manifest neutralizing antibodies, suggesting that SCoV-specific neutralizing antibodies are important for the suppression of viral replication within the lungsORGAN.
displaCy
DespiteINsomeDTdifferencesNNSinINtheDTcellularJJcompositionNNofINinflammatoryJJinfiltrates,NNSwePRPdidVBDnotRBobserveVBanyDTovertJJlungNNpathologyNNinINtheDTchimeric-VLPtreatedJJmice,NNSwhenWRBcomparedVBNtoTOtheDTnegativeJJcontrolNNmice.NNScasedetnmodcasedetamodnmodcaseamodnmodnsubjauxnegdetamodcompounddobjcasedetamodnmodadvmodadvclcasedetamodcompoundnmod
displaCy
Despite some differences in the cellularCELL composition of inflammatory infiltrates, we did not observe any overt lungORGAN pathology in the chimeric-VLPtreated miceORGANISM, when compared to the negative control miceORGANISM.
displaCy
OurPRP$resultsNNSshowVBPthatINchimericJJVLPNNcanMDbeVBanDTeffectiveJJvaccineNNstrategyNNagainstINSCoVNNinfection.NNcc:preconjnsubjmarkamodnsubjauxpredetdetamodcompoundccompcaseamodnmod
displaCy
Our results show that chimeric VLPCELL can be an effective vaccine strategy against SCoVSIMPLE_CHEMICAL infection.
displaCy
"``SevereJJacuteJJrespiratoryJJsyndrome (NNSARS)NNisVBZaDTnewlyRBemergedVBNdiseaseNNcausedVBNbyINSARSNNPcoronavirus (NNSCoV).NNpunctamodamodamodnsubjappospredetdetadvmodamodaclcasecompoundnmodappos
displaCy
"Severe acute respiratory syndrome (SARS) is a newly emerged disease caused by SARS coronavirusORGANISM (SCoVSIMPLE_CHEMICAL).
displaCy
SARSNNPoriginatedVBDinINSouthernNNPChinaNNPinIN2002CDandCCspreadVBDtoTOfiveCDdifferentJJcontinentsNNScausingVBG>SYM8000CDinfectionNNandCC>XX700CDdeathsNNSbeforeINitsPRP$apparentJJeradicationNNasINaDThumanJJinfectionNNinIN2004 .CDnsubjprepcompoundpobjpreppobjccconjprepnummodamodpobjaclpunctnummoddobjccpunctnummodconjcasecc:preconjamodnmodcasedetamodnmodcasenmod
displaCy
SARS originated in Southern China in 2002 and spread to five different continents causing >8000 infection and >700 deaths before its apparent eradication as a humanORGANISM infection in 2004 .
displaCy
ItPRPisVBZnotRBknownVBNifINtheDTvirusNNwillMDbeVBreintroducedVBNintoINtheDThumanJJpopulationNNbutCCancestralJJcoronavirusesNNSareVBPwidelyRBdistributedVBNinINbatsNNSandCCareVBPthoughtVBNtoTOhaveVBadaptedVBNtoINcivetsNNSandCCthenRBtoTOhumansNNSinINrecentJJtimeNNperiods [NNS2,CD3] .CDnsubjpassauxpassnegmarkdetnsubjpassauxauxpassadvclcasedetamodnmodccamodnsubjpassauxpassadvmodconjcasenmodccauxpassconjauxauxxcomppreppobjccadvmodpreppobjprepamodcompoundpobjconjnsubj
displaCy
It is not known if the virus will be reintroduced into the humanORGANISM population but ancestral coronavirusesORGANISM are widely distributed in batsORGANISM_SUBSTANCE and are thought to have adapted to civetsORGANISM and then to humansORGANISM in recent time periods [2, 3] .
displaCy
BecauseINemergingVBGvirusesNNStendVBPtoTOreemergeVBasINconditionsNNSchange ,VBPitPRPisVBZhighlyRBdesirableJJtoTOdevelopVBsafeJJandCCefficaciousJJvaccinesNNSand/orCCantiviralsNNStoTOpreventVBSCoVNNinfections.NNSmarkamodnsubjadvclmarkxcompmarkcompoundnmodnsubjpredetadvmodmarkxcompamodccconjdobjccconjmarkxcompamoddobj
displaCy
Because emerging viruses tend to reemerge as conditions change , it is highly desirable to develop safe and efficacious vaccines and/or antivirals to prevent SCoVSIMPLE_CHEMICAL infections.
displaCy
"``AllDTcoronaviruses,NNSincludingVBGSCoV,NNcarryVBPfourCDstructuralJJproteins:NNSnucleocapsid (NNN)NNproteinNNandCCthreeCDenvelopeNNproteins,NNSnamelyRBspike (JJS)NNprotein,NNaDTtypeNNICDtransmembraneNNglycoprotein;NNenvelope (NNE)NNprotein;NNandCCmembrane (NNM)NNprotein,NNwhichWDThasVBZthreeCDmembrane-spanningJJdomains.NNSpunctdetnsubjcasenmodnummodamoddobjcompoundcompoundagentccnummodcompoundconjadvmodamodcompoundapposdetcompoundnummodcompoundapposcompoundcompoundconjcccompoundcompoundconjnsubjparataxisnummodamoddobj
displaCy
"All coronavirusesORGANISM, including SCoVSIMPLE_CHEMICAL, carry four structural proteins: nucleocapsidGENE_OR_GENE_PRODUCT (N) protein and three envelope proteins, namely spike (S) protein, a type I transmembrane glycoproteinGENE_OR_GENE_PRODUCT; envelope (E) protein; and membrane (M) proteinGENE_OR_GENE_PRODUCT, which has three membrane-spanning domains.
displaCy
CoronavirusNNSNNproteinNNisVBZresponsibleJJforINvirusNNadsorptionNNtoTOsusceptibleJJcellsNNSthroughINaDTspecificJJvirus-receptorJJinteractionNNandCCinducesVBZmembraneNNfusionNNbetweenINviralJJenvelopeNNandCChostNNcellNNmembrane .NNcompoundcompoundnsubjpredetcasecompoundnmodcaseamodnmodcasedetamodcompoundnmodccconjcompounddobjcaseamodnmodcccompoundcompoundconj
displaCy
Coronavirus SORGANISM protein is responsible for virus adsorption to susceptible cellsCELL through a specific virus-receptor interaction and induces membraneCELLULAR_COMPONENT fusion between viral envelope and host cell membrane .CELL
displaCy
SNNproteinNNisVBZaDTmainJJplayerNNforINdeterminingVBGcoronavirusNNtissueNNtropism,NNhostNNspecificityNNandCCviralJJpathogenicity .NNcompoundnsubjpredetdetamodmarkaclcompoundcompounddobjcompoundconjccamodconj
displaCy
S protein is a main player for determining coronavirus tissueORGANISM tropism, host specificity and viral pathogenicity .
displaCy
BecauseINmostJJScoronavirusNNneutralizingNNantibodiesNNSrecognizeVBPSNNprotein [NN1,CD13] ,CDitPRPisVBZnotRBsurprisingJJthatINmostJJSofINtheDTcurrentJJSCoVNNvaccineNNcandidatesNNSareVBPeitherCCtheDTSNNproteinNNsubunitNNitselfPRPorCCthoseDTcarryingVBGSNNprotein .NNmarkamodcompoundcompoundnsubjadvclcompounddobjagentnsubjnsubjpredetnegmarknsubjprepdetamodamodcompoundpobjpredetnmod:npmoddetcompoundcompoundccompagentccconjaclcompounddobj
displaCy
Because most coronavirusORGANISM neutralizing antibodies recognize S protein [1, 13] , it is not surprising that most of the current SCoV vaccineGENE_OR_GENE_PRODUCT candidates are either the S protein subunit itself or those carrying S protein .
displaCy
Furthermore,RBprophylacticJJadministrationNNofINmonoclonalJJantibodiesNNSdirectedVBNatINtheDTSCoVNNSNNproteinNNprotectsVBZanimalsNNSagainstINsubsequentJJSCoVNNchallenge .NNadvmodamodnsubjcaseamodnmodaclcasedetamodcompoundnmoddobjcaseamodamodnmod
displaCy
Furthermore, prophylactic administration of monoclonal antibodies directed at the SCoV SGENE_OR_GENE_PRODUCT protein protects animals against subsequent SCoVSIMPLE_CHEMICAL challenge .
displaCy
TheseDTstudiesNNSpointVBPoutRPthatINneutralizingVBGantibodiesNNSthatWDTrecognizeVBPSCoVNNSNNproteinNNareVBPsufficientJJtoTOpreventVBorCCdecreaseVBtheDTmorbidityNNandCCmortalityNNassociatedVBNwithINSCoVNNinfectionNNbyINprimarilyRBsuppressingVBGreplicationNNofINtheDTchallengeNNvirus.NNdetnsubjprtmarkamodnsubjnsubjparataxisamodcompounddobjpredetccompmarkxcompccconjdetdobjccconjaclcaseamodnmodmarkadvmodadvcldobjcasedetcompoundnmod
displaCy
These studies point out that neutralizing antibodies that recognize SCoV SGENE_OR_GENE_PRODUCT protein are sufficient to prevent or decrease the morbidity and mortality associated with SCoVSIMPLE_CHEMICAL infection by primarily suppressing replication of the challenge virus.
displaCy
"``Coronavirus-likeJJparticles (NNSVLPs)NNSareVBPproducedVBNfromINtheDTcellsNNScoexpressingVBGtheDTS,NNM,NNandCCENNproteins ;NNSexpressionNNofINtheDTlatterJJtwoCDproteinsNNSareVBPsufficientJJforINVLPNNproduction .NNpunctamodnsubjpassapposauxpasscasedetnmodacldetdobjconjcccompoundconjnsubjcasedetamodnummodnmodpredetmwecasecompoundnmod
displaCy
"Coronavirus-like particlesGENE_OR_GENE_PRODUCT (VLPsORGANISM) are produced from the cellsCELL coexpressing the S, M, and E proteins ; expression of the latter two proteins are sufficient for VLPCELL production .
displaCy
MNNproteinNNplaysVBZaDTcentralJJroleNNinINvirusNNassembly,NNwhileINSNNproteinNNisVBZassembledVBNintoINcoronavirusNNparticlesNNSthroughINSNNprotein-MJJproteinNNinteraction .NNcompoundnsubjdetamoddobjcasecompoundnmodmarkcompoundnsubjpassauxpassadvclcasecompoundnmodcasecompoundcompoundcompoundnmod
displaCy
M protein plays a central role in virus assembly, while S protein is assembled into coronavirus particles throughORGANISM S protein-M protein interaction .
displaCy
Further,RBinteractionsNNSofINtheDTMNNproteinNNwithINtheDTRNANNpackagingNNsignalNNofINtheDTviralJJRNANNandCCwithINNNNproteinNNdriveVBPincorporationNNofINtheDThelicalJJnucleocapsidNNcomplex,NNwhichWDTconsistsVBZofINtheDTviralJJgenomeNNandCCNNNprotein,NNintoINvirusNNparticles.NNSadvmodnsubjcasedetcompoundnmodcasedetcompoundcompoundnmodcasedetamodnmodcccasecompoundconjdobjcasedetamodcompoundnmodnsubjparataxiscasedetamodnmodcccompoundconjcasecompoundnmod
displaCy
Further, interactions of the MGENE_OR_GENE_PRODUCT protein with the RNA packaging signal of the viral RNA and with N protein drive incorporation of the helical nucleocapsid complex, which consists of the viral genome and N protein, into virus particlesORGANISM.
displaCy
VacciniaNNvirusNNand/orCCalphavirusNNrepliconsNNShaveVBPbeenVBNusedVBNtoTOexpressVBcoronavirusNNproteinsNNStoTOenableVBgenerationNNofINVLPs ,NNSwhileINwePRPhaveVBPreportedVBNproductionNNofINSCoVNNVLPsNNSfromIN293CDTNNcellsNNSthatWDTareVBPco-transfectedVBNwithINfourCDeukaryoticJJpCAGGS-basedJJexpressionNNplasmids,NNSeachDTofINwhichWDTencodesVBZSCoVNNS,NNM,NNNNNandCCENNproteins .NNScompoundnsubjpasscccompoundconjauxauxpassmarkxcompcompounddobjmarkxcompdobjcasenmodmarknsubjauxadvcldobjcaseamodnmodcasenummodcompoundnmodnsubjpassauxpassparataxiscasenummodamodamodcompoundnmodnsubjcasenmodadvclamodcompoundconjconjccconjdobj
displaCy
Vaccinia virusORGANISM and/or alphavirusORGANISM replicons have been used to express coronavirusORGANISM proteins to enable generation of VLPsCELL , while we have reported production of SCoV VLPsORGANISM from 293T cellsCELL that are co-transfected with four eukaryotic pCAGGS-based expression plasmids, each of which encodes SCoV SGENE_OR_GENE_PRODUCT, MGENE_OR_GENE_PRODUCT, N and E proteins .
displaCy
OthersNNShaveVBPalsoRBreportedVBNproductionNNofINSCoVNNVLPNNfromINinsectJJcells [NNS37,CD38]CDandCCmammalianJJcells .NNSnsubjauxadvmoddobjcaseamodnmodcaseamodnmodagentconjccamodconj
displaCy
Others have also reported production of SCoV VLPGENE_OR_GENE_PRODUCT from insect cellsCELL [37, 38] and mammalian cellsCELL .
displaCy
"''DuringINourPRP$studiesNNSofINcoronavirusNNassembly,NNwePRPfoundVBDanDTefficientJJproductionNNofINchimericJJVLPsNNScarryingVBGSCoVNNSNNproteinNNandCCmurineJJcoronavirus (NNmouseNNhepatitisNNvirusNNorCCMHV)NNM,NNNNNandCCENNproteinsNNSfromINcellsNNScoexpressingVBGthoseDTproteins.NNSpunctcasecc:preconjnmodcasecompoundnmodnsubjdetamoddobjcaseamodnmodaclamodcompounddobjccamodconjcompoundcompoundcompoundccconjcompoundconjccconjnmodcasenmodacldetdobj
displaCy
"During our studies of coronavirusORGANISM assembly, we found an efficient production of chimeric VLPsCELL carrying SCoV SGENE_OR_GENE_PRODUCT protein and murine coronavirusORGANISM (mouse hepatitis virusORGANISM or MHV) M, N and E proteins from cellsCELL coexpressing those proteins.
displaCy
InINmiceNNSimmunizedVBNwithINtheDTchimericJJVLPs,NNStheDTpresentJJstudyNNdescribesVBZelicitationNNofINantibodiesNNSthatWDTneutralizedVBDSCoVNNandCCsuppressedVBDchallengedVBNSCoVNNreplicationNNinINtheDTlungs.NNScasenmodaclcasedetamodnmoddetamodnsubjdobjcasenmodnsubjparataxisdobjccconjamodamoddobjcasedetnmod
displaCy
In miceORGANISM immunized with the chimeric VLPsCELL, the present study describes elicitation of antibodies that neutralized SCoVSIMPLE_CHEMICAL and suppressed challenged SCoVSIMPLE_CHEMICAL replication in the lungsORGAN.
displaCy
TheseDTfindingsNNSsuggestVBPthatINtheDTuseNNofINchimericJJVLPNNisVBZanDTeffectiveJJvaccineNNstrategyNNagainstINSCoVNNinfection.NNdetnsubjmarkdetnsubjcaseamodnmodpredetdetamodcompoundccompcaseamodnmod
displaCy
These findings suggest that the use of chimeric VLPCELL is an effective vaccine strategy against SCoVSIMPLE_CHEMICAL infection.
displaCy
"``VeroNNE6NNcells,NNS293CDTNNcellsNNSandCCCHONNcellsNNSwereVBDgrownVBNinINDulbeccoNNP'sPOSmodifiedVBNminimumJJessentialJJmedium (NNDMEM)NNsupplementedVBNwithINpenicillin (NN100CDunits/ml),NNSstreptomycin (NN100CDg/ml),NN0.2%CDsodiumNNbicarbonateNNandCC10%CDfetalJJbovineJJserum (NNFBS).NNpunctcompoundcompoundnmodnummodcompoundnsubjpasscccompoundconjauxpasscasecc:preconjcaseamodamodamodnmodapposaclcasenmodnummodapposconjnummodapposconjcompoundagentccamodamodamodconjappos
displaCy
"Vero E6 cellsCELL, 293T cellsCELL and CHO cellsCELL were grown in Dulbecco'sORGANISM modified minimum essential medium (DMEMSIMPLE_CHEMICAL) supplemented with penicillinSIMPLE_CHEMICAL (100 units/ml), streptomycinSIMPLE_CHEMICAL (100 g/mlSIMPLE_CHEMICAL), 0.2% sodium bicarbonateSIMPLE_CHEMICAL and 10% fetal bovine serum (FBSORGANISM_SUBSTANCE).
displaCy
TheDTUrbaniNNstrainNNofINSCoVNNwasVBDobtainedVBNfromINT.G.NNPKsiazekNNPatINtheDTCentersNNPSforINDiseaseNNPControlNNPandCCPrevention (NNPAtlanta,NNPGA),NNPandCCaDTworkingVBGstockNNofINthisDTvirusNNwasVBDpreparedVBNbyINseriallyRBpassagingVBGaDTportionNNofINtheDTseedNNvirusNNtwiceRBinINVeroNNE6NNcells.NNSdetcompoundnsubjpasscasenmodauxpasscasecompoundnmodprepdetpobjprepcompoundpobjccconjagentapposccdetamodnsubjpasscasedetnmodauxpassconjmarkadvmodadvcldetdobjprepdetcompoundpobjadvmodcasecompoundcompoundnmod
displaCy
The Urbani strain of SCoVSIMPLE_CHEMICAL was obtained from T.G. Ksiazek at the Centers for Disease Control and Prevention (Atlanta, GAGENE_OR_GENE_PRODUCT), and a working stockORGANISM of this virus was prepared by serially passaging a portion of the seed virus twice in Vero E6 cellsCELL.
displaCy
TheDTcultureNNfluidNNfromINinfectedJJcellsNNSwasVBDclarifiedVBNbyINlow-speedJJcentrifugationNNandCCwasVBDfilteredVBNusingVBGaDT0.45CDmNNfilter,NNportioned,VBNandCCfrozenVBNatIN\u221280NNP\u2022NNPC.NNPMediumNNPfromINuninfectedJJVeroNNE6NNcellsNNSwasVBDsimilarlyRBtreatedVBNandCCprocessed.VBNdetcompoundnsubjpasscaseamodnmodauxpasscaseamodnmodccauxpassconjxcompdetnummodcompounddobjconjccconjprepcompoundcompoundcompoundpobjcaseamodcompoundcompoundnmodauxpassadvmodconjccconj
displaCy
The culture fluid from infected cellsCELL was clarified by low-speed centrifugation and was filtered using a 0.45 m filter, portioned, and frozen at \u221280 \u2022 C. Medium from uninfected Vero E6 cellsCELL was similarly treated and processed.
displaCy
"``TheDTplasmids,NNSpCAGGS-MHV-A59/S,NNpCAGGS-MHV-A59/E,NNpCAGGS-MHV-A59/M,NNandCCpCAGGS-A59/N,NNwereVBDconstructedVBNbyINinsertingVBGtheDTentireJJregionNNofINMHVNNstructuralJJproteins,NNSS,NNE,NNPMNNandCCN,NNrespectively,RBintoINaDTchickenNNbeta-actinJJpromoterbasedJJexpressionNNplasmid,NNpCAGGS-MCS;NNplasmidNNpMH54NNwasVBDobtainedVBNfromINPaulNNPMastersNNPandCCusedVBNasINaDTtemplateNNforINPCRNNamplificationNNofINMHVNNstructuralJJgenes.NNSpunctdetnsubjpassconjconjconjccconjauxpassmarkadvcldetamoddobjcasecompoundamodnmodapposapposapposccconjadvmodcasedetcompoundcompoundamodcompoundnmodapposcompoundnsubjpassauxpassparataxisprepcompoundpobjccconjcasedetnmodcasecompoundnmodcasecompoundamodnmod
displaCy
"The plasmids, pCAGGS-MHV-A59/SGENE_OR_GENE_PRODUCT, pCAGGS-MHV-A59/E, pCAGGS-MHV-A59/MCELL, and pCAGGS-A59/NSIMPLE_CHEMICAL, were constructed by inserting the entire region of MHVORGANISM structural proteins, S, E, M and N, respectively, into a chicken beta-actinORGANISM promoterbased expression plasmid, pCAGGS-MCS; plasmid pMH54 was obtained from Paul Masters and used as a template for PCR amplification of MHVORGANISM structural genes.
displaCy
ConstructionNNofINplasmids,NNSpCAGGS-SNNexpressingVBGSCoVNNSNNprotein,NNpCAGGS-ENNexpressingVBGSCoVNNENNprotein,NNpCAGGS-MNNexpressingVBGSCoVNNMNNprotein,NNandCCpCAGGS-NNNexpressingVBGSCoVNNNNNprotein,NNwasVBDdescribedVBNpreviously .RBnsubjpasscasenmodnsubjpassaclamodcompounddobjapposaclamodcompounddobjapposaclamodcompounddobjccconjaclamodcompounddobjauxpassadvmod
displaCy
Construction of plasmids, pCAGGS-SSIMPLE_CHEMICAL expressing SCoV SGENE_OR_GENE_PRODUCT protein, pCAGGS-EGENE_OR_GENE_PRODUCT expressing SCoV EGENE_OR_GENE_PRODUCT protein, pCAGGS-MGENE_OR_GENE_PRODUCT expressing SCoV MGENE_OR_GENE_PRODUCT protein, and pCAGGS-N expressing SCoV NGENE_OR_GENE_PRODUCT protein, was described previously .
displaCy
"''ForINpreparationNNofINMHVNNVLPs,NNSsubconfluentJJ293CDTNNcellNNculturesNNSinIN100-mmJJtissueNNcultureNNdishesNNSwereVBDcotransfectedVBNwithIN14CDgNNofINpCAGGS-MHV-A59/S,NN0.4CDgNNofINpCAGGS-MHV-A59/E,NN3.5CDgNNofINpCAGGS-MHV-A59/M,NNandCC14CDgNNofINpCAGGS-A59/NNNusingVBGTransIT-293NNreagent (NNMirus).NNpunctpreppobjcasecompoundnmodamodnummodcompoundcompoundnsubjpasscaseamodcompoundcompoundnmodauxpasscasenummodnmodcasenmodnummodapposcasenmodnummodapposcasenmodccnummodconjcasenmodaclcompounddobjappos
displaCy
"For preparation of MHV VLPsORGANISM, subconfluent 293T cell culturesCELL in 100-mm tissueTISSUE culture dishes were cotransfected with 14 g of pCAGGS-MHV-A59/SGENE_OR_GENE_PRODUCT, 0.4 g of pCAGGS-MHV-A59/E, 3.5 g of pCAGGS-MHV-A59/MCELL, and 14 g of pCAGGS-A59/N using TransIT-293GENE_OR_GENE_PRODUCT reagent (Mirus).
displaCy
AtIN3CDdaysNNSposttransfection,NNtheDTcultureNNmediaNNSwereVBDcollected,VBNcentrifugedVBNatIN1550CD\u00d7NNSgNNforIN10CDminNNandCCfilteredVBNthroughINaDT0.45-mNNfilterNNtoTOremoveVBcellNNdebris.NNcasecompoundamodnmoddetcompoundnsubjpassauxpassadvclcasenummodamodnmodcasenummodnmodccconjprepdetcompoundpobjauxxcompcompounddobj
displaCy
At 3 days posttransfection, the culture media were collected, centrifuged at 1550 \u00d7 g for 10 min and filtered through a 0.45-m filter to remove cellCELL debris.
displaCy
TheDTMHVNNVLPsNNSwereVBDpelletedVBNdownRPthroughIN20%CDsucroseNNcushionNNatIN26,000CDrpmNNforIN3CDhNNbyINusingVBGaDTBeckmanNNPSWNNP28CDrotor.NNdetcompoundnsubjpassauxpassadvmodprepcompoundcompoundpobjcasenummodnmodcasenummodnmodmarkadvcldetcompoundcompoundnummoddobj
displaCy
The MHV VLPsORGANISM were pelleted down through 20% sucroseSIMPLE_CHEMICAL cushion at 26,000 rpm for 3 h by using a Beckman SW 28 rotor.
displaCy
AfterINtheDTpelletsNNSwereVBDsuspendedVBNinINNTENNbuffer (NN100CDmMNNNaCl,NN10CDmMNNTris-HCl,NNpHNN7.0,CD1CDmMNNEDTA),NNtheDTVLPsNNSwereVBDcentrifugedVBNusingVBGaDTBeckmanNNPSWNNP28CDrotorNNatIN26,000CDrpmNNforIN3CDhNNonINaDTdiscontinuousJJsucroseNNgradientNNconsistingVBGofIN60,CD50,CD30,CDandCC20%CDsucrose.NNmarkdetnsubjpassauxpassadvclcasecompoundnmodcompoundamodagentcompoundamodconjapposconjcompoundamodapposdetnsubjpassauxpassxcompdetcompoundcompoundnummoddobjcasenummodnmodcasenummodnmodcasedetamodcompoundnmodaclcasenummodconjconjcccompoundnmod
displaCy
After the pellets were suspended in NTE bufferSIMPLE_CHEMICAL (100 mM NaCl, 10 mM Tris-HCl, pH 7.0, 1 mM EDTA), the VLPsORGANISM_SUBSTANCE were centrifuged using a Beckman SW 28 rotor at 26,000 rpm for 3 h on a discontinuous sucroseSIMPLE_CHEMICAL gradient consisting of 60, 50, 30, and 20% sucroseSIMPLE_CHEMICAL.
displaCy
TheDTVLPsNNSatINtheDTinterfaceNNofIN30CDandCC50%CDsucroseNNwereVBDcollected,VBNdilutedVBNandCCfurtherRBpurifiedVBNonINaDTdiscontinuousJJsucroseNNgradientNNconsistingVBGofIN60,CD50,CD30,CDandCC20%CDsucroseNNatIN26,000CDrpmNNforIN18CDh.NNPurifiedVBNandCCconcentratedVBNVLPsNNSatINtheDTinterfaceNNbetweenIN50CDandCC30%CDsucroseNNwereVBDcollected,VBNdilutedVBNandCCpelletedVBNthroughINaDT20%CDsucroseNNcushionNNatIN26,000CDrpmNNforIN2CDh.NNTheDTpelletsNNSwereVBDsuspendedVBNinINNTENNbufferNNandCCkeptVBDatIN\u221280NNP\u2022NNPCNNuntilINfurtherJJuse.NNdetnsubjpassprepdetpobjprepcompoundcccompoundpobjauxpassconjccadvmodconjcasedetamodcompoundnmodaclcasenummodconjconjcccompoundnmodcasenummodnmodcasenummodamodnmodccamodconjcasedetnmodcasecompoundcccompoundnmodauxpassconjconjccconjcasedetcompoundcompoundnmodcasenummodnmodcasenummodnmoddetnsubjpassauxpassconjcasecompoundnmodccconjcasecompoundcompoundnmodcaseamodnmod
displaCy
The VLPsCELL at the interface of 30 and 50% sucroseSIMPLE_CHEMICAL were collected, diluted and further purified on a discontinuous sucroseSIMPLE_CHEMICAL gradient consisting of 60, 50, 30, and 20% sucroseSIMPLE_CHEMICAL at 26,000 rpm for 18 h. Purified and concentrated VLPsCELL at the interface between 50 and 30% sucroseSIMPLE_CHEMICAL were collected, diluted and pelleted through a 20% sucroseSIMPLE_CHEMICAL cushion at 26,000 rpm for 2 h. The pellets were suspended in NTE bufferSIMPLE_CHEMICAL and kept at \u221280 \u2022 CGENE_OR_GENE_PRODUCT until further use.
displaCy
ForINpreparationNNofINSCoVNNVLP,NNaDTmixtureNNofIN2.8CDgNNofINpCAGGS-S,NN13.2CDgNNofINpCAGGS-E,NN1.4CDgNNofINpCAGGS-M,NNandCC2.8CDgNNofINpCAGGS-NNNwasVBDcotransfectedVBNintoIN293CDTNNcellsNNSgrownVBNonINaDT100-mmJJtissueNNcultureNNplate.NNcasenmodcaseamodnmoddetapposcasenummodnmodcasenmodnummodapposcasenmodnummodconjcasenmodccnummodnsubjpasscasenmodauxpasscasenummodcompoundnmodaclcasedetamodcompoundcompoundnmod
displaCy
For preparation of SCoV VLPCELL, a mixture of 2.8 g of pCAGGS-S, 13.2 g of pCAGGS-EGENE_OR_GENE_PRODUCT, 1.4 g of pCAGGS-MGENE_OR_GENE_PRODUCT, and 2.8 g of pCAGGS-N was cotransfected into 293T cellsCELL grown on a 100-mm tissueTISSUE culture plate.
displaCy
ChimericJJVLPsNNSwereVBDgeneratedVBNbyINtransfectingVBG293CDTNNcellsNNSorCCCHONNcellsNNSinINaDT100-mmJJtissueNNcultureNNplateNNwithIN14CDgNNofINpCAGGS-S,NN0.4CDgNNofINpCAGGS-MHV-A59/E,NN3.5CDgNNofINpCAGGS-MHV-A59/M,NNandCC14CDgNNofINpCAGGS-A59/N.NNSCoVNNVLPsNNSandCCchimericJJVLPsNNSwereVBDpurifiedVBNusingVBGtheDTsameJJprocedureNNasINdescribedVBNforINtheDTMHVNNVLPNNpurification.NNamodnsubjpassauxpassmarkadvclnummodcompounddobjcccompoundconjcasedetamodcompoundcompoundnmodcasenummodnmodcasenmodnummodapposcasenmodnummodapposcasenmodccnummodconjcasenmodamodagentccamodconjauxpassconjxcompdetamoddobjmarkadvclcasedetcompoundcompoundnmod
displaCy
Chimeric VLPsORGANISM were generated by transfecting 293T cellsCELL or CHO cellsCELL in a 100-mm tissueTISSUE culture plate with 14 g of pCAGGS-S, 0.4 g of pCAGGS-MHV-A59/E, 3.5 g of pCAGGS-MHV-A59/MCELL, and 14 g of pCAGGS-A59/N. SCoV VLPs and chimeric VLPs were purified using the same procedure as described for the MHV VLPORGANISM purification.
displaCy
"``TheDTtotalJJproteinNNconcentrationNNofINVLPsNNSwasVBDquantitatedVBNbyINBio-RadNNDCNNproteinNNassayNNaccordingVBGtoINtheDTmanufacturerNN'sPOSinstructions (NNSBIO-RAD,NNCA).NNpunctdetamodcompoundnsubjpasscasenmodauxpasscasecompoundcompoundcompoundnmodprepquantmoddetposscasepobjapposappos
displaCy
"The total protein concentration of VLPsORGANISM was quantitated by Bio-Rad DC protein assay according to the manufacturer's instructions (BIO-RAD, CA).
displaCy
ForINdetectionNNofINMHVNNS,NNMNNandCCNNNproteins,NNSwePRPusedVBDanti-MHVNNserum,NNwhichWDTwasVBDproducedVBNbyINimmunizingVBGrabbitsNNSwithINaDTpurifiedVBNJHMNNstrainNNofINMHV,NNkindlyRBprovidedVBNbyINSusanNNPBaker,NNPLoyolaNNPUniversityNNPofINChicago.NNPcasenmodcasecompoundnmodconjccconjagentnsubjamoddobjnsubjpassauxpassparataxismarkadvcldobjcasedetamodcompoundnmodcasenmodadvmodadvcldativecompoundpobjcompoundappospreppobj
displaCy
For detection of MHV S,ORGANISM MGENE_OR_GENE_PRODUCT and N proteins, we used anti-MHV serum, which was produced by immunizing rabbitsORGANISM with a purified JHMORGANISM strain of MHVORGANISM, kindly provided by Susan Baker, Loyola University of Chicago.
displaCy
ForINdetectionNNofINSCoVNNSNNprotein,NNaDTmixtureNNofINpurifiedVBNrabbitNNpolyclonalJJanti-SCoVNNSNNproteinNNantibody (NNABGENT,NNcat.NNcasecaseamodcompoundnmoddetapposcaseamodcompoundamodcompoundcompoundcompoundnmodagentappos
displaCy
For detection of SCoV SGENE_OR_GENE_PRODUCT protein, a mixture of purified rabbit polyclonal anti-SCoV S protein antibodyORGANISM (ABGENT, catORGANISM.
displaCy
no.NNAP6000a)NNandCCpolyclonalJJrabbitNNanti-SCoVNNSNNproteinNNantibody (NNIMGENEX,NNcat.NNcompoundccamodcompoundcompoundcompoundcompoundconjapposappos
displaCy
no. AP6000aORGANISM) and polyclonal rabbit anti-SCoV S protein antibodyORGANISM (IMGENEXGENE_OR_GENE_PRODUCT, catORGANISM.
displaCy
SCoVNNNNNproteinNNwasVBDdetectedVBNbyINusingVBGrabbitNNpolyclonalJJanti-SCoVNNNNNproteinNNantibody (NNIMGENEX,NNcat.NNamodcompoundnsubjpassauxpassmarkadvclcompoundamodcompoundcompoundcompounddobjagentappos
displaCy
SCoV NSIMPLE_CHEMICAL protein was detected by using rabbit polyclonal anti-SCoV N protein antibodyORGANISM (IMGENEXGENE_OR_GENE_PRODUCT, catORGANISM.
displaCy
no.NNIMGNN548),CDandCCSCoVNNMNNproteinNNwasVBDdetectedVBNbyINusingVBGaDTmixtureNNofINSCoVNNPUPMNNantibody-N-terminal (NNABGENT,NNcat.NNcompoundnsubjpassnsubjccamodcompoundconjauxpassmarkadvcldetdobjcaseamodpreconjagentnmodappos
displaCy
no. IMG 548), and SCoV MGENE_OR_GENE_PRODUCT protein was detected by using a mixture of SCoV PUPMGENE_OR_GENE_PRODUCT antibody-N-terminal (ABGENT, catORGANISM.
displaCy
no.JJAP6008a),NNSCoVNNPUPMNNantibodyNNC-terminal (JJABGENT,NNcat.NNcompoundapposcompoundpreconjxcompagentappos
displaCy
no. AP6008aORGANISM), SCoV PUPM antibodyGENE_OR_GENE_PRODUCT C-terminal (ABGENT, catORGANISM.
displaCy
no.NNAP6008b)NNandCCanti-SCoVNNMNNantibodies (NNSProSci,NNcat.NNcompoundccamodcompoundconjapposappos
displaCy
no. AP6008b) and anti-SCoV M antibodiesGENE_OR_GENE_PRODUCT (ProSciGENE_OR_GENE_PRODUCT, catORGANISM.
displaCy
no.NN3527PNNPandCCcatNNno.NN3529P).NNPamodcccompoundconjappos
displaCy
no. 3527P and cat no. 3529PORGANISM).
displaCy
"''AfterINwashingVBGtheDTgelNNwithINwater,NNproteinsNNSinINtheDTgelNNwereVBDstainedVBNbyINsoakingVBGtheDTgelNNwithINBio-SafeNNCoomassieNNStain (NNBIO-RAD)NNwithINgentleJJagitationNNforIN1CDh.NNTheDTgelNNwasVBDrinsedVBNextensivelyRBwithINwaterNNovernight.RBpunctpreppcompdetdobjpreppobjnsubjpasscasedetnmodauxpassadvclpreppcompdetdobjprepcompoundcompoundpobjapposprepamodpobjprepnummodpobjdetnsubjpassauxpassadvmodpreppobjadvmod
displaCy
"After washing the gel with water, proteins in the gel were stained by soaking the gel with Bio-Safe Coomassie Stain (BIO-RAD)SIMPLE_CHEMICAL with gentle agitation for 1 h. The gel was rinsed extensively with water overnight.
displaCy
"``Carbon-coated,JJ200-meshJJcopperNNgridsNNSwereVBDfloatedVBNonINdropsNNSofINchimericJJVLPNNforIN10CDmin.NNpunctamodamodcompoundnsubjpassauxpasspreppobjcaseamodnmodcasenummodnmod
displaCy
"Carbon-coated, 200-mesh copper grids were floated on drops of chimeric VLPORGANISM_SUBSTANCE for 10 min.
displaCy
AfterINwashingVBGtheDTgridsNNSwithINwaterNNthreeCDtimes,NNSnegativeJJstainingNNwasVBDperformedVBNusingVBG2%CDphosphotungsticJJacid (NNpHNN7.0)CDforIN1CDmin.NNpreppcompdetdobjpreppobjnummodnpadvmodamodnsubjpassauxpassxcompnummodamoddobjapposnsubjcasenummodnmod
displaCy
After washing the grids with water three times, negative staining was performed using 2% phosphotungstic acidSIMPLE_CHEMICAL (pH 7.0) for 1 min.
displaCy
"''section": "NNElectronNNmicroscopicJJanalysisNNofINchimericJJVLP"NN_SPpunctcompoundamodagentprepamodpunct
displaCy
"section": "ElectronCELLULAR_COMPONENT microscopic analysis of chimeric VLPCELL"
displaCy
"''Six-toJJ8-week-old,JJfemaleJJBalb/cNNmice (NNSCharlesNNPRiverNNPlaboratory,NNWilmington,NNPMA)NNPwereVBDhousedVBNinINcagesNNScoveredVBNwithINbarrierNNfiltersNNSinINanDTapprovalNNbiosafetyNNlevelNN3CDanimalNNfacilityNNmaintainedVBNbyINtheDTUniversityNNPofINTexasNNPMedicalNNPBranchNNPatINGalveston,NNPTexas.NNPpunctnsubjpassagentamodcompoundconjcompoundcompoundapposconjapposauxpasspreppobjaclprepcompoundpobjprepdetcompoundcompoundcompoundnummodcompoundpobjacldativedetpobjprepcompoundcompoundpobjpreppobjappos
displaCy
"Six-to 8-week-oldORGANISM, female Balb/c miceORGANISM (Charles River laboratory, Wilmington, MAGENE_OR_GENE_PRODUCT) were housed in cages covered with barrier filters in an approval biosafety level 3 animal facility maintained by the University of Texas Medical Branch at Galveston, Texas.
displaCy
AllDTofINtheDTmouseNNexperimentsNNSwereVBDperformedVBNusingVBGexperimentalJJprotocolsNNSapprovedVBNbyINtheDTUniversityNNPofINTexasNNPMedicalNNPBranchNNPInvestigationalNNPAnimalNNPCareNNPandCCUseNNPCommittee;NNPallDTofINtheDTexperimentsNNSwereVBDcarriedVBNoutRPfollowingVBGNationalNNPInstitutesNNPSofINHealthNNPandCCUnitedNNPStatesNNPDepartmentNNPofINAgricultureNNPguidelines.NNSnsubjpasscasedetcompoundnmodauxpassxcompamoddobjacldativedetpobjprepcompoundcompoundcompoundcompoundcompoundpobjcccompoundconjnsubjpasscasedetnmodauxpassmweprtcasecompoundnmodpreppobjcccompoundcompoundconjprepcompoundpobj
displaCy
All of the mouseORGANISM experiments were performed using experimental protocols approved by the University of Texas Medical Branch Investigational Animal Care and Use Committee; all of the experiments were carried out following National Institutes of Health and United States Department of Agriculture guidelines.
displaCy
"``MHVNNVLPs,NNSchimericJJVLPsNNSorCCinfluenzaNNvirusNNvaccineNNwereVBDadjustedVBNtoTOtwiceRBtheirPRP$desiredJJfinalJJconcentration (NNg/0.1NNml)NNandCCthenRBmixedVBNwithINanDTequalJJvolumeNNofINphosphateNNbufferedJJsaline (NNPBS)NNorCCalumNNadjuvant (NNImjectNNalum,NNPierce,NNcat.NNpunctcompoundnsubjpassamodnsubjpasscccompoundcompoundconjauxpassprepadvmodcc:preconjamodamodnmodcompoundapposccadvmodconjcasedetamodnmodcasecompoundamodnmodapposcccompoundconjamodagentapposappos
displaCy
"MHV VLPsORGANISM, chimeric VLPsORGANISM or influenza virus vaccineORGANISM were adjusted to twice their desired final concentration (g/0.1 ml) and then mixed with an equal volume of phosphate buffered salineSIMPLE_CHEMICAL (PBS) or alumSIMPLE_CHEMICAL adjuvant (Imject alum, PierceGENE_OR_GENE_PRODUCT, catORGANISM.
displaCy
AtINdayNN0,CDtheyPRPwereVBDadministeredVBNintramuscularly (RBi.m.)NNusingVBG1-mlJJtuberculinNNsyringes (NNSNorm-Ject,NNTuttingen,NNPGermany)NNPandCC26-g,NN3/8-in.NNcasenmodnsubjnsubjpassauxpassadvmodagentxcompamodcompounddobjagentconjapposccconjconj
displaCy
At day 0, they were administered intramuscularly (i.m.) using 1-ml tuberculin syringes (Norm-JectGENE_OR_GENE_PRODUCT, Tuttingen, Germany) and 26-g, 3/8-in.
displaCy
beveledVBNneedles (NNSBectonNNPDickinson,NNPcat.NNamodcompoundapposappos
displaCy
beveled needles (Becton Dickinson, catORGANISM.
displaCy
InINcontrolJJgroups,NNSmiceNNSwereVBDinoculatedVBNi.m.NNcasecompoundnmodnsubjpassauxpassdobj
displaCy
In control groups, miceORGANISM were inoculated i.m.
displaCy
withINVeroNNE6NNcellNNcultureNNfluid,NNaDTmixtureNNofINinfluenzaNNvaccineNNandCCalum,NNorCCalumNNalone,RBorCCtheyPRPwereVBDleftVBNuntreated.JJcasecompoundcompoundcompoundcompounddetapposcasecompoundnmodccconjccconjadvmodccnsubjpassauxpassconjxcomp
displaCy
with Vero E6 cellCELL culture fluid, a mixture of influenza vaccine and alumSIMPLE_CHEMICAL, or alumSIMPLE_CHEMICAL alone, or they were leftORGAN untreated.
displaCy
MiceNNSwereVBDinoculatedVBNonINdayNN0CDand,CCasINindicatedVBNinINtheDTbodyNNofINtheDTpaperNNaDTsecondJJinjectionNNwasVBDgivenVBNonINdayNN28CDinINmostJJSexperiments.NNSnsubjpassauxpasscasenmodnsubjccmarkadvclcasedetnmodprepdetpobjdetamodnsubjpassauxpassconjcasenmodnsubjcaseamodnmod
displaCy
MiceORGANISM were inoculated on day 0 and, as indicated in the bodyORGANISM_SUBDIVISION of the paper a second injection was given on day 28 in most experiments.
displaCy
SCoVNNwasVBDinoculatedVBNdirectlyRBintoINtheDTnaresNNSofINmice (NNS40CDl)NNthatWDTwereVBDlightlyRBanesthetizedVBNwithINisoflurane (NNIsoFlo;NNAbbottNNPLaboratories;NNPSNorthNNPChicago,NNPIL).NNPnsubjpassauxpassadvmodcasedetnmodcasenmodnummodapposnsubjpassauxpassadvmodparataxiscasenmodapposcompoundapposcompoundapposappos
displaCy
SCoVSIMPLE_CHEMICAL was inoculated directly into the naresORGAN of miceORGANISM (40 l) that were lightly anesthetized with isofluraneSIMPLE_CHEMICAL (IsoFlo; Abbott Laboratories; North Chicago, ILGENE_OR_GENE_PRODUCT).
displaCy
MiceNNSthatWDTwereVBDinoculatedVBNwithIN1CD\u00d7NN10CD6CDTCIDNN50CDofINSCoVNNatINdayNN0CDtoTOprovideVBpost-infectionJJimmunityNNdidVBDnotRBundergoVBaDTsecondJJinoculation.NNnsubjnsubjpassauxpassparataxiscasenummodexplnummodnummodnmodnsubjcasenmodcasenmodnsubjmarknmod:possamoddobjauxnegdetamoddobj
displaCy
MiceORGANISM that were inoculated with 1 \u00d7 10 6 TCID 50 of SCoVSIMPLE_CHEMICAL at day 0 to provide post-infection immunity did not undergo a second inoculation.
displaCy
InINmostJJSofINourPRP$experiments,NNSimmunizedJJmiceNNSwereVBDchallengedVBNbyINintranasal (JJi.n.)NNSinoculationNNofIN1CD\u00d7NN10CD6CDTCIDNN50CDofINSCoVNNatINdayNN56CDandCCtheyPRPwereVBDeuthanizedVBNatINdayNN58,CDwhileINinINsomeDTexperimentsNNSmiceNNSwereVBDinoculatedVBNwithINSCoVNNatINdayNN28CDandCCtheyPRPwereVBDeuthanizedVBNatINdayNN30.CDcasenmodcasecc:preconjnmodamodnsubjpassauxpasscasenmodapposnmodcasenummodexplnummodnummodnmodnsubjcasenmodcasenmodnsubjccnsubjpassauxpassconjcasenmodnsubjmarkcasedetcompoundnsubjpassauxpassadvclcasenmodcasenmodnsubjccnsubjpassauxpassconjcasenmodnsubj
displaCy
In most of our experiments, immunized miceORGANISM were challenged by intranasal (i.n.) inoculation of 1 \u00d7 10 6 TCID 50 of SCoVSIMPLE_CHEMICAL at day 56 and they were euthanized at day 58, while in some experiments miceORGANISM were inoculated with SCoVSIMPLE_CHEMICAL at day 28 and they were euthanized at day 30.
displaCy
"``ProceduresNNSforINimmunizationNNandCCSCoVNNchallenge"NN_SPpunctpreppobjccamodpunct
displaCy
"Procedures for immunization and SCoVSIMPLE_CHEMICAL challenge"
displaCy
"''MiceNNSwereVBDanesthetizedVBNwithINisofluraneNNandCCthenRBbledVBNfromINtheDTretro-orbitalJJsinusJJplexus.NNpunctnsubjpassauxpasscasenmodccadvmodconjcasedetamodamodnmod
displaCy
"MiceORGANISM were anesthetized with isofluraneSIMPLE_CHEMICAL and then bled from the retro-orbital sinusMULTI-TISSUE_STRUCTURE plexus.
displaCy
AfterINheatNNinactivationNNatIN56CD\u2022NNCNNforIN30CDmin,NNseraNNSwereVBDstoredVBNatIN4CD\u2022NNPC.NNPTheDTassayNNforINvirusspecificJJneutralizingVBGantibodiesNNSwasVBDperformedVBNonINserialJJ2-foldRBdilutedVBNsamplesNNSofINeachDTserumNNbeginningVBGatINaDTdilutionNNofIN1:5CDusingVBG2%CDFBS-DMEMNNasINtheDTdiluentNNinIN96-wellJJtissueNNcultureNNplates (NNSFalconNNP3072);CDtheDTfinalJJvolumeNNofINtheDTseriallyRBdilutedVBNsamplesNNSinINeachDTwellNNwasVBD60CDl.NNAfterINadditionNNofIN120CDTCIDNN50CDofINSCoVNNinIN60CDlNNintoINeachDTwell,RBtheDTsamplesNNSwereVBDincubatedVBNforIN45-CD60CDminNNatINroomNNtemperature.NNcasecompoundnmodcasenummodcompoundnmodcasenummodnmodnsubjpassauxpasscasenummodcompoundcompounddetnmodcaseamodamodnmodauxpassconjcaseamodagentamodnmodcasedetnmodaclcasedetnmodcasenmodaclamoddobjcasedetnmodcasecompoundcompoundcompoundnmodapposnsubjdetamodnsubjcasedetadvmodamodnmodcasedetnmodpredetnummodmwecasenmodcasenummodnmodnsubjcasenmodcasenummodnmodcasedetnmoddetnsubjpassauxpassmwecasecompoundnummodnmodcasecompoundnmod
displaCy
After heat inactivation at 56 \u2022 C for 30 min, seraORGANISM_SUBSTANCE were stored at 4 \u2022 C. The assay for virusspecific neutralizing antibodies was performed on serial 2-fold diluted samples of each serumORGANISM_SUBSTANCE beginning at a dilution of 1:5 using 2% FBS-DMEMORGANISM_SUBSTANCE as the diluent in 96-well tissueTISSUE culture plates (Falcon 3072); the final volume of the serially diluted samples in each well was 60 l.CELL After addition of 120 TCID 50 of SCoVSIMPLE_CHEMICAL in 60 l into each well, the samples were incubated for 45-60 min at room temperature.
displaCy
ThenRB100CDlNNofINtheseDTmixtures,NNScontainingVBG100CDTCIDNN50CDofINSCoV,NNwereVBDtransferredVBNintoINduplicateJJwellsNNSofINconfluentJJVeroNNE6NNcellsNNSgrownVBNinIN96-wellJJmicrotiterNNplates.NNSadvmodnummodnsubjpasscasedetnmodaclnummoddobjnsubjcasenmodauxpasscaseamodnmodcaseamodcompoundcompoundnmodaclcasecompoundcompoundnmod
displaCy
Then 100 l of these mixtures, containing 100 TCID 50 of SCoVSIMPLE_CHEMICAL, were transferred into duplicate wells of confluent Vero E6 cellsCELL grown in 96-well microtiter plates.
displaCy
AfterIN72CDhNNincubation,NNwhenWRBtheDTvirusNNcontrolNNwellsNNSexhibitedVBDadvancedJJvirus-inducedJJCPE,NNtheDTneutralizingVBGcapacityNNofINindividualJJserumNNsamplesNNSwasVBDassessedVBNbyINdeterminingVBGtheDTpresenceNNorCCabsenceNNofINvirusinducedJJCPE.NNcasenummodcompoundnmodadvmoddetcompoundcompoundnsubjadvclamodamoddobjdetamodnsubjpasscaseamodcompoundnmodauxpassmarkadvcldetdobjccconjcaseamodnmod
displaCy
After 72 h incubation, when the virus control wells exhibited advanced virus-induced CPE, the neutralizing capacity of individual serum samplesORGANISM_SUBSTANCE was assessed by determining the presence or absence of virusinduced CPECELL.
displaCy
SCoV-specificJJneutralizingVBGantibodyNNtitersNNSwereVBDexpressedVBNasINtheDTreciprocalNNofINtheDTlastJJdilutionNNofINserumNNthatWDTcompletelyRBinhibitedVBDvirus-inducedJJCPE.NNamodamodcompoundnsubjpassauxpasscasedetnmodcasedetamodnmodcasenmodnsubjadvmodparataxisamoddobj
displaCy
SCoV-specific neutralizing antibody titers were expressed as the reciprocal of the last dilution of serumORGANISM_SUBSTANCE that completely inhibited virus-induced CPE.
displaCy
"``TwoCDdaysNNSpostVBPSCoVNNchallenge,NNmiceNNSwereVBDeuthanizedVBNandCCtheirPRP$lungsNNSwereVBDremoved.VBNpunctnummodnpadvmodadvclamoddobjnsubjpassauxpasscccc:preconjnsubjpassauxpassconj
displaCy
"Two days post SCoVSIMPLE_CHEMICAL challenge, miceORGANISM were euthanized and their lungsORGAN were removed.
displaCy
LungNNlobes,NNSincludingVBGrightJJmiddleJJlobe,NNrightRBlowerJJRlobe,NNaccessoryNNlobeNNandCCleftVBDlobe,NNwereVBDremovedVBNandCCplacedVBNinIN10%CDbufferedJJformalinNNforINsubsequentJJhistologicalJJexaminationNNandCCimmunohistochemistry (NNIHC)NNasINdescribedVBNpreviously .RBcompoundnsubjpasscaseamodamodnmodadvmodamodconjamodconjccamodconjauxpassccconjcasecompoundamodnmodcaseamodamodnmodccconjapposmarkadvcladvmod
displaCy
Lung lobesMULTI-TISSUE_STRUCTURE, including right middle lobeCANCER, right lower lobeCANCER, accessory lobeCANCER and left lobeCANCER, were removed and placed in 10% buffered formalin for subsequent histological examination and immunohistochemistry (IHC) as described previously .
displaCy
ForINvirusNNtitration,NNtheDTremainingVBGlobesNNSofINtheDTlungsNNSwereVBDweighedVBNandCCfrozenVBNatIN\u221280NN\u2022NNCNNbeforeINbeingVBGhomogenizedVBNinINPBS/10%JJFBSNNsolutionNNusingVBGtheDTTissueLyser (NNPQiagen;NNPRetsch,NNPHaan,NNPGermany).NNPcasecompoundnmoddetamodnsubjpasscasedetnmodauxpassccconjcasecompoundcompoundnmodmarkauxpassadvclcaseamodcompoundnmodacldetdobjapposapposconjappos
displaCy
For virus titration, the remaining lobesORGAN of the lungsORGAN were weighed and frozen at \u221280 \u2022 C before being homogenized in PBS/10% FBSORGANISM_SUBSTANCE solution using the TissueLyserCANCER (Qiagen; Retsch, Haan, Germany).
displaCy
TheDThomogenatesNNSwereVBDthenRBcentrifuged,VBNandCCSCoVNNtitersNNSinINtheDTclarifiedVBNfluidNNwereVBDdeterminedVBNinINaDTTCIDNN50CDassayNNinINquadruplicateJJwellsNNSofINVeroNNE6NNcellsNNSinIN96-wellJJplates.NNSdetnsubjpassauxpassadvmodccamodnsubjpasscasedetamodnmodauxpassconjcasedetcompoundnummodnmodcaseamodnmodcasecompoundcompoundnmodcasecompoundnmod
displaCy
The homogenatesORGANISM were then centrifuged, and SCoVSIMPLE_CHEMICAL titers in the clarified fluid were determined in a TCIDSIMPLE_CHEMICAL 50 assay in quadruplicate wells of Vero E6 cellsCELL in 96-well plates.
displaCy
TitersNNSofINvirusNNinINlungNNhomogenatesNNSwereVBDexpressedVBNasINTCIDNN50CD/gramNNofINtheDTlungs (NNSlogNN10 )CDandCCtheDTminimalJJdetectableJJlevelNNofINvirusNNwasVBD2.3CDlogNN10CDTCIDNN50CD/g.NNSnsubjpasscasenmodcasecompoundnmodauxpasscasecompoundnummodnmodcasedetnmodagentnsubjccdetamodamodconjcasenmodpredetnummodcompoundnummodnmodnsubjpunct
displaCy
Titers of virus in lung homogenatesCELL were expressed as TCIDSIMPLE_CHEMICAL 50 /gram of the lungsORGAN (log 10 ) and the minimal detectable level of virus was 2.3 log 10 TCID 50 /g.
displaCy
"''section": "NNCollectionNNofINlungs,NNShistology,NNimmunohistochemistry,NNandCCvirusNNtitration"NN_SPpunctconjcasenmodconjconjcccompoundpunct
displaCy
"section": "Collection of lungsORGAN, histology, immunohistochemistry, and virus titration"
displaCy
"''Formalin-fixedJJandCCparaffin-embeddedJJtissueNNsectionsNNSwereVBDsubjectedVBNtoTOtheDTstandardJJhematoxylinNNandCCeosinNNandCCIHCNNstainingNNforINevaluatingVBGhistopathologyNNandCCdetectingVBGSCoVNNantigen,NNrespectively .RBpunctamodccconjcompoundnsubjpassauxpasscasedetamodnmodccconjcccompoundconjmarkadvcldobjccconjamoddobjadvmod
displaCy
"Formalin-fixedORGAN and paraffin-embedded tissue sectionsTISSUE were subjected to the standard hematoxylin and eosin and IHCGENE_OR_GENE_PRODUCT staining for evaluating histopathology and detecting SCoV antigenGENE_OR_GENE_PRODUCT, respectively .
displaCy
"''section": "NNCollectionNNofINlungs,NNShistology,NNimmunohistochemistry,NNandCCvirusNNtitration"NN_SPpunctconjcasenmodconjconjcccompoundpunct
displaCy
"section": "Collection of lungsORGAN, histology, immunohistochemistry, and virus titration"
displaCy
"''ByINusingVBGaDTstatisticalJJprogramNNInstat, (NNPversionNN3,CDGraphpadNNPSoftware,NNPInc.,NNPSanNNPDiego,NNPCA),NNwePRPperformedVBDtheDTKruskal-WallisNNnon-parametricJJanalysisNNofINvariance (NNANOVA)NNtestNNtoTOcompareVBarithmeticJJmeanNNSCoVNNlungNNtitersNNSandCCneutralizingVBGantibodyNNtiters;NNSundetectableJJvirusNNtitersNNSwereVBDassignedVBNaDTvalueNNofIN1.8 (CDtheDTminimalJJdetectionNNlimitNNbeingVBG2.3CDlogNN10CD/gNNlung)NNandCCundetectableJJvirus-specificJJneutralizingVBGtitersNNSwereVBDassignedVBNaDTvalueNNofIN10.CDpunctpreppcompdetamodcompounddobjnpadvmodnsubjcompoundconjconjcompoundconjapposnsubjadvcldetcompoundamoddobjcasecompoundapposnmodmarkaclamodamodamodcompounddobjccamodcompoundconjamodcompoundnsubjpassauxpassmwedetdobjcasenmoddetamodcompoundagentpredetnummodcompoundnsubjagentnmodccamodamodamodconjauxpassdetdobjpreppobj
displaCy
"By using a statistical program Instat, (version 3, Graphpad Software, Inc., San Diego, CA), we performed the Kruskal-Wallis non-parametric analysis of variance (ANOVA) test to compare arithmetic mean SCoV lungORGAN titers and neutralizing antibody titers; undetectable virus titers were assigned a value of 1.8 (the minimal detection limit being 2.3 log 10 /g lungORGAN) and undetectable virus-specificGENE_OR_GENE_PRODUCT neutralizing titers were assigned a value of 10.
displaCy
"``WePRPhaveVBPpreviouslyRBreportedVBNtheDTsuccessfulJJproductionNNofINSCoVNNVLPsNNSfromIN293CDTNNcellsNNScotransfectedVBNwithINfourCDpCAGGSbasedJJplasmids,NNSeachDTofINwhichWDTencodesVBZSCoVNNS,NNM,NNNNNorCCENNproteinsNNSwithoutINusingVBGexogenousJJvirusesNNSforINproteinNNexpression.NNpunctnsubjauxadvmoddetamoddobjcaseamodnmodcasenummodcompoundnmodaclcasenummodamodnmodnsubjcasenmodmweamodcompoundconjconjccconjdobjmarkadvclamoddobjcasecompoundnmod
displaCy
"We have previously reported the successful production of SCoV VLPsCELL from 293T cellsCELL cotransfected with four pCAGGSbasedGENE_OR_GENE_PRODUCT plasmids, each of which encodes SCoV SGENE_OR_GENE_PRODUCT, MGENE_OR_GENE_PRODUCT, N or E proteins without using exogenous viruses for protein expression.
displaCy
InINtheDTstudyNNreportedVBNhereRBwePRPvariedVBDtheDTamountNNofINeachDTplasmidNNforINcotransfectionNNandCCwereVBDableJJtoTOgenerateVBapproximatelyRB1.3CDgNNofINSCoVNNVLPsNNSfromIN2CD\u00d7NNS10CD7CD293CDTNNcells.NNScasedetnmodacladvmodnsubjdetdobjcasedetnmodcasenmodccpredetconjmarkxcompadvmodnummoddobjcaseamodnmodcasecompoundadvmodcompoundcompoundnummodcompoundnmod
displaCy
In the study reported here we varied the amount of each plasmid for cotransfection and were able to generate approximately 1.3 g of SCoV VLPsORGANISM from 2 \u00d7 10 7 293T cellsSIMPLE_CHEMICAL.
displaCy
ColloidalNNCoomassieNNblueJJstainingNNandCCWesternNNblotNNanalysisNNidentifiedVBDSCoVNNS,NNNNNandCCMNNproteinsNNSinINtheDTpurifiedVBNSCoVNNVLPs (NNSFig.NN1A) .NNcompoundcompoundamodnsubjcccompoundcompoundconjamoddobjconjccconjagentcasedetamodamodnmodcompoundappos
displaCy
Colloidal Coomassie blueGENE_OR_GENE_PRODUCT staining and Western blot analysis identified SCoV SGENE_OR_GENE_PRODUCT, N and MGENE_OR_GENE_PRODUCT proteins in the purified SCoV VLPsCELL (Fig. 1ACELL) .
displaCy
Likewise,RBtransfectionNNofINfourCDpCAGGSbasedJJplasmids,NNSeachDTofINwhichWDTencodedVBDMHVNNS,NNPE,NNMNNorCCNNNproteins,NNSresultedVBDinINproductionNNofINMHVNNVLPs (NNSFig.NN1A) .NNadvmodnsubjcasenummodamodnmodnsubjcasenmodparataxiscompoundcompoundconjconjccconjdobjcasenmodcasecompoundnmodcompoundappos
displaCy
Likewise, transfection of four pCAGGSbased plasmidsGENE_OR_GENE_PRODUCT, each of which encoded MHV S,ORGANISM E, M or N proteins, resulted in production of MHV VLPsORGANISM (Fig. 1A) .
displaCy
AfterINoptimization,NNwePRPobtainedVBDapproximatelyRB22.3CDgNNofINpurifiedVBNMHVNNVLPsNNSfromIN2CD\u00d7NNS10CD7CD293CDTNNcells,NNSdemonstratingVBGthatINtheDTefficiencyNNofINMHVNNVLPNNproductionNNwasVBDsubstantiallyRBbetterJJRthanINthatDTofINSCoVNNVLPNNproduction.NNcasenmodnsubjadvmodnummoddobjcaseamodcompoundnmodcasenummodamodnummodnummodagentcompoundnmodadvclmarkdetnsubjcasecompoundcompoundnmodpredetadvmodccompcasenmodcaseamodcompoundnmod
displaCy
After optimization, we obtained approximately 22.3 g of purified MHV VLPsORGANISM from 2 \u00d7 10 7 293T cells, demonstrating that the efficiency of MHV VLPORGANISM production was substantially better than that of SCoV VLPORGANISM production.
displaCy
BecauseINMNNandCCENNproteinsNNSdriveVBPVLPNNassemblyNNandCCrelease ,NNandCCcoronavirusNNSNNproteinNNisVBZincorporatedVBNintoINVLPsNNSthroughINM-SNNinteraction [NN25,CD27,CD28,CD42] ,CDtheseDTdataNNSindicatedVBDthatINMHVNNMNNandCCENNproteinsNNSresultedVBDinINmoreRBRefficientJJVLPNNproductionNNthanINdidVBDSCoVNNMNNandCCENNproteins.NNSmarkcompoundccconjnsubjadvclcompounddobjccconjcccompoundcompoundnsubjpassauxpassconjcasenmodcaseamodnmodagentconjconjapposdetnsubjmarkcompoundcompoundcccompoundnsubjccompcaseadvmodamodcompoundnmodexpladvclamodcompoundcccompounddobj
displaCy
Because M and E proteins drive VLPCELL assembly and release , and coronavirus S proteinORGANISM is incorporated into VLPsCELL through M-S interaction [25, 27, 28, 42] , these data indicated that MHV MORGANISM and EGENE_OR_GENE_PRODUCT proteins resulted in more efficient VLPORGANISM_SUBSTANCE production than did SCoV MGENE_OR_GENE_PRODUCT and EGENE_OR_GENE_PRODUCT proteins.
displaCy
"``ProductionNNandCCcharacterizationNNofINchimericJJVLPs"NNS_SPpunctccconjcaseamodnmod
displaCy
"Production and characterization of chimeric VLPsCELL"
displaCy
"``WePRPnextRBtestedVBDwhetherINcoexpressionNNofINSCoVNNSNNproteinNNandCCMHVNNE,NNNNNandCCMNNproteinNNcouldMDresultVBinINtheDTefficientJJproductionNNofINchimericJJVLPsNNSwhichWDTwouldMDcontainVBhighJJconcentrationsNNSofINtheDTSCoVNNSNNprotein.NNpunctnsubjadvmodmarknsubjcaseamodcompoundnmodcccompoundcompoundapposccconjconjauxccompcasedetamodnmodcaseamodnmodnsubjauxparataxisamoddobjcasedetamodcompoundnmod
displaCy
"We next tested whether coexpression of SCoV SGENE_OR_GENE_PRODUCT protein and MHV EORGANISM, N and MGENE_OR_GENE_PRODUCT protein could result in the efficient production of chimeric VLPsORGANISM_SUBSTANCE which would contain high concentrations of the SCoV SGENE_OR_GENE_PRODUCT protein.
displaCy
IfINSCoVNNSNNproteinNNinteractsVBZwithINMHVNNMNNprotein,NNthenRBtheDTrobustJJVLPNNproductionNNmachineryNNdrivenVBNbyINMHVNNMNNandCCENNproteins,NNScouldMDresultVBinINefficientJJchimericJJVLPNNproduction.NNmarkamodcompoundnsubjadvclcasecompoundcompoundnmodadvmoddetamodcompoundcompounddobjaclcasecompoundnmodcccompoundconjauxcaseamodamodcompoundnmod
displaCy
If SCoV SGENE_OR_GENE_PRODUCT protein interacts with MHV MORGANISM protein, then the robust VLPCELL production machinery driven by MHV MORGANISM and EGENE_OR_GENE_PRODUCT proteins, could result in efficient chimeric VLPCELL production.
displaCy
ChimericJJVLPNNproduction,NNindeed,RBoccurredVBDfromIN293CDTNNcellsNNSthatWDTwereVBDcotransfectedVBNwithINplasmids,NNSeachDTofINwhichWDTexpressedVBNSCoVNNSNNprotein,NNMHVNNNNNprotein,NNMHVNNMNNproteinNNorCCMHVNNENNprotein (NNFig.NN1A) .NNamodcompoundnsubjadvmodcasenummodcompoundnmodnsubjpassauxpassparataxiscasenmodnsubjcasenmodmweamodcompounddobjcompoundcompoundconjcompoundcompoundconjcccompoundcompoundconjcompoundappos
displaCy
Chimeric VLPCELL production, indeed, occurred from 293T cellsCELL that were cotransfected with plasmids, each of which expressed SCoV SGENE_OR_GENE_PRODUCT protein, MHV N proteinORGANISM, MHV M protein or MHV E protein (Fig. 1A) .
displaCy
ColloidalNNCoomassieNNblueJJstainingNNandCCWesternNNblotNNanalysisNNofINpurifiedVBNchimericJJVLPNNshowedVBDtheDTpresenceNNofINSCoVNNS,NNMHVNNNNNandCCMHVNNMNNproteinsNNSinINchimericJJVLPs,NNSwithINonlyRBaDTlowJJlevelNNofINhostNNproteinNNcontamination.NNcompoundcompoundamodnsubjcccompoundcompoundconjcaseamodamodnmoddetdobjcaseamodnmodcompoundcompoundcccompoundconjapposcaseamodnmodcaseadvmoddetamodnmodcasecompoundcompoundnmod
displaCy
Colloidal Coomassie blueGENE_OR_GENE_PRODUCT staining and Western blot analysis of purified chimeric VLPCELL showed the presence of SCoV SGENE_OR_GENE_PRODUCT, MHV NORGANISM and MHV M proteins in chimeric VLPsCELL, with only a low level of host protein contamination.
displaCy
NegativeJJstainingNNofINchimericJJVLPNNrevealedVBDaDTtypicalJJsphericalJJcoronavirusNNstructureNNwithINpeplomers (NNSFig.NN1B) .NNamodnsubjcaseamodnmoddetamodamodcompounddobjcasenmodcompoundappos
displaCy
Negative staining of chimeric VLPCELL revealed a typical spherical coronavirusORGANISM structure with peplomers (Fig. 1B) .
displaCy
AfterINoptimization,NNapproximatelyRB8.7CDgNNofINpurifiedVBNchimericJJVLPsNNSwasVBDproducedVBNfromIN2CD\u00d7NNS10CD7CD293CDTNNcells;NNStheDTefficiencyNNofINchimericJJVLPNNproductionNNwasVBDaboutRB8CDtimesNNSbetterJJRthanINthatDTofINSCoVNNVLPs.NNScasenmodadvmodnummodnsubjpasscaseamodamodnmodauxpasscasenummodadvmodcompoundcompoundnummodcompoundnmoddetnsubjcaseamodcompoundnmodpredetadvmodnummodnpadvmodmwecasenmodcaseamodnmod
displaCy
After optimization, approximately 8.7 g of purified chimeric VLPsCELL was produced from 2 \u00d7 10 7 293T cells; the efficiency of chimeric VLPCELL production was about 8 times better than that of SCoV VLPsORGANISM.
displaCy
"``ProductionNNandCCcharacterizationNNofINchimericJJVLPs"NNS_SPpunctccconjcaseamodnmod
displaCy
"Production and characterization of chimeric VLPsCELL"
displaCy
"``WePRPnextRBtestedVBNefficienciesNNSofINchimericJJVLPNNproductionNNfromINVeroNNandCCCHONNcells,NNSbothDTofINwhichWDThaveVBPbeenVBNusedVBNforINtheDTpreparationNNofINhumanJJvaccines .NNSpunctdetadvmodamodnsubjpasscaseamodcompoundnmodcasecompoundccconjnmodnsubjpasscasenmodauxauxpasscasedetnmodcaseamodnmod
displaCy
"We next tested efficiencies of chimeric VLPCELL production from VeroCELL and CHO cellsCELL, both of which have been used for the preparation of human vaccinesORGANISM .
displaCy
ByINusingVBGvariousJJDNANNtransfectionNNreagentsNNSandCCcombinationsNNSofINdifferentJJconcentrationsNNSofINeachDTplasmidNNforINtransfection,NNtheDTefficienciesNNSofINchimericJJVLPNNproductionNNfromINVeroNNcellsNNSwereVBDaboutRB10CDtimesNNSlowerJJRthanINthoseDTfromIN293CDTNNcells (NNSdataNNSnotRBshown).VBNmarkadvclamodcompoundcompounddobjccconjcaseamodnmodcasedetnmodcasenmoddetnsubjcaseamodcompoundnmodcasecompoundnmodpredetadvmodnummodnpadvmodcasenmodcasenummodcompoundnmodagentnegacl
displaCy
By using various DNACELLULAR_COMPONENT transfection reagents and combinations of different concentrations of each plasmid for transfection, the efficiencies of chimeric VLPCELL production from Vero cellsCELL were about 10 times lower than those from 293T cellsCELL (data not shown).
displaCy
WePRPfoundVBDthatINwhenWRBwePRPusedVBDTransIT-293,NNPtheDTproductionNNofINchimericJJVLPsNNSfromINCHONNcellsNNSwasVBDaboutRBone-thirdCDofINthoseDTfromIN293CDTNNcellsNNSinINrepeatedVBNexperiments (NNSdataNNSnotRBshown).VBNnsubjmarkadvmodnsubjadvcldobjdetnsubjcaseamodnmodcasecompoundnmodpredetadvmodccompcasenmodcasenummodcompoundnmodcaseamodnmodagentnegacl
displaCy
We found that when we used TransIT-293GENE_OR_GENE_PRODUCT, the production of chimeric VLPsCELL from CHO cellsCELL was about one-third of those from 293T cellsCELL in repeated experiments (data not shown).
displaCy
ProlongedJJincubationNNofINtransfectedVBN293CDTNNcellsNNSandCCCHONNcellsNNSforINmoreJJRthanIN4CDdaysNNSposttransfectionNNdidVBDnotRBimproveVBtheDTaccumulationNNofINchimericJJVLPsNNSinINtheDTsupernatant (JJdataNNSnotRBshown).VBNamodnsubjcaseamodnummodcompoundnmodcccompoundconjcaseadvmodquantmodnummodcompoundnmodauxnegdetdobjcaseamodnmodcasedetnmodagentnegacl
displaCy
Prolonged incubation of transfected 293T cellsCELL and CHO cellsCELL for more than 4 days posttransfection did not improve the accumulation of chimeric VLPsCELL in the supernatant (data not shown).
displaCy
"``ProductionNNandCCcharacterizationNNofINchimericJJVLPs"NNS_SPpunctccconjcaseamodnmod
displaCy
"Production and characterization of chimeric VLPsCELL"
displaCy
"''ForINassessingVBGtheDTusefulnessNNofINchimericJJVLPsNNSasINaDTSCoVNNvaccineNNcandidate,NNwePRPfirstRBtestedVBDtheDTeffectsNNSofINimmunizationNNofINmiceNNSwithINchimericJJVLPsNNSonINserumNNneutralizingVBGantibodyNNproductionNNandCCinhibitionNNofINSCoVNNreplicationNNinINtheDTlung.NNpunctpreppcompdetdobjcaseamodnmodcasedetamodcompoundnmodnsubjadvmoddetdobjcasenmodcasenmodcaseamodnmodcasecompoundamodcompoundnmodccconjcaseamodnmodcasedetnmod
displaCy
"For assessing the usefulness of chimeric VLPsCELL as a SCoV vaccineSIMPLE_CHEMICAL candidate, we first tested the effects of immunization of miceORGANISM with chimeric VLPs on serumORGANISM_SUBSTANCE neutralizing antibody production and inhibition of SCoVSIMPLE_CHEMICAL replication in the lungORGAN.
displaCy
SixtoNN8-week-oldJJfemaleJJBalb/cNNmiceNNSwereVBDinoculatedVBNi.m.NNamodamodamodcompoundnsubjpassauxpassdobj
displaCy
Sixto 8-week-oldORGANISM female Balb/c miceORGANISM were inoculated i.m.
displaCy
onceRBwithINaDTmixtureNNofIN2CDgNNofINchimericJJVLPsNNSpreparedVBNfromIN293CDTNNwithINplacebo (NNmediumNNfromINVeroNNE6NNcells),NNSaDTmixtureNNofIN2CDgNNchimericJJVLPNNandCCPBSNNorCCaDTmixtureNNofIN2CDgNNchimericJJVLPNNandCCalumNNorCCi.n.NNSadvmodcasedetcasenummodnmodcaseamodnmodaclcasenummodnmodcasenmodagentcasecompoundcompoundnmoddetapposcasenummodnmodamodagentccconjccdetconjcasenummodnmodamodagentccconjccconj
displaCy
once with a mixture of 2 g of chimeric VLPsCELL prepared from 293TORGANISM with placebo (medium from Vero E6 cellsCELL), a mixture of 2 g chimeric VLPCELL and PBS or a mixture of 2 g chimeric VLPCELL and alumSIMPLE_CHEMICAL or i.nORGANISM_SUBSTANCE.
displaCy
withIN1CD\u00d7NN10CD6CDTCIDNN50CDofINinfectiousJJSCoV.NNAfterIN28CDdays,NNStheseDTmiceNNSwereVBDchallengedVBNwithIN1CD\u00d7NN10CD6CDTCIDNN50CDofINSCoV.NNTwoCDdaysNNSlater,RBRmiceNNSwereVBDsacrificedVBNandCCtheDTvirusNNtitersNNSinINtheDTlungsNNSwereVBDdeterminedVBNasINshownVBNinINtheDTgraph.NNcasenummodexplnummodnummodnmodnsubjcaseamodnmodcasenummodnmoddetnsubjpassauxpasscasenummodexplnummodnummodnmodnsubjcasenmodnummodnpadvmodadvmodnsubjpassauxpassconjccdetcompoundnsubjpasscasedetnmodauxpassconjmarkadvclcasedetnmod
displaCy
with 1 \u00d7 10 6 TCID 50 of infectious SCoV.GENE_OR_GENE_PRODUCT After 28 days, these miceORGANISM were challenged with 1 \u00d7 10 6 TCID 50 of SCoV.GENE_OR_GENE_PRODUCT Two days later, miceORGANISM were sacrificed and the virus titers in the lungsORGAN were determined as shown in the graph.
displaCy
cellsNNSandCCPBSNNorCC2CDgNNofINchimericJJVLPNNmixedVBNwithINalum.NNccconjccnummodconjcaseamodnmodaclcasenmod
displaCy
cellsCELL and PBS or 2 g of chimeric VLPCELL mixed with alumSIMPLE_CHEMICAL.
displaCy
AsINaDTplacebo,NNmiceNNSwereVBDinoculatedVBNi.m.NNcasedetnmodnsubjpassauxpassdobj
displaCy
As a placebo, miceORGANISM were inoculated i.m.
displaCy
withINVeroNNE6NNcellNNcultureNNfluid,NNwhileINanotherDTgroupNNofINmiceNNSwereVBDinoculatedVBNi.n.NNScasecompoundcompoundcompoundcompoundnmodmarkdetnsubjpasscasenmodauxpassdobj
displaCy
with Vero E6 cellCELL culture fluid, while another group of miceORGANISM were inoculated i.n.
displaCy
withIN1CD\u00d7NN10CD6CDTCIDNN50CDofINSCoVNNtoTOproduceVBsolidJJpost-infectionJJimmunity.NNcasenummodexplnummodnummodnsubjcasenmodmarkaclamodamoddobj
displaCy
with 1 \u00d7 10 6 TCID 50 of SCoVSIMPLE_CHEMICAL to produce solidCANCER post-infection immunity.
displaCy
Twenty-eightCDdaysNNSlater,RBtheDTmiceNNSwereVBDbledVBNforINdeterminingVBGtheDTserumNNneutralizingNNantibodyNNtiters (NNSFig.NN2A) .NNnummodnpadvmodadvmoddetnsubjpassauxpassmarkadvcldetcompoundcompoundcompounddobjcompoundappos
displaCy
Twenty-eight days later, the miceORGANISM were bled for determining the serumORGANISM_SUBSTANCE neutralizing antibody titers ( Fig. 2A) .
displaCy
Subsequently,RBtheDTmiceNNSwereVBDchallengedVBNi.n.NNSadvmoddetnsubjpassauxpassdobj
displaCy
Subsequently, the miceORGANISM were challenged i.nORGANISM.
displaCy
withIN1CD\u00d7NN10CD6CDTCIDNN50CDofINSCoVNNandCCtheDTlungNNvirusNNtitersNNSatIN2CDdaysNNSpostVBPSCoVNNchallengeNNwereVBDdetermined (VBNFig.NN2B) .NNcasenummodexplnummodnummodnmodnsubjcasenmodccdetcompoundcompoundconjcasenummodnmodamodnsubjpassauxpassccompcompoundagent
displaCy
with 1 \u00d7 10 6 TCID 50 of SCoVSIMPLE_CHEMICAL and the lung virusMULTI-TISSUE_STRUCTURE titers at 2 days post SCoVSIMPLE_CHEMICAL challenge were determined (Fig. 2B) .
displaCy
NoDTSCoV-specificJJneutralizingVBGantibodiesNNSwereVBDdetectableJJinINtheDTseraNNofINtheDTmiceNNSinoculatedJJi.m.NNnegamodamodnsubjpredetcasedetnmodcasedetnmodacldobj
displaCy
No SCoV-specific neutralizing antibodies were detectable in the seraORGANISM_SUBSTANCE of the miceORGANISM inoculated i.m.
displaCy
MinimalJJorCCnoDTvirus-specificJJneutralizingVBGantibodiesNNSwereVBDevidentJJinINmiceNNSinoculatedVBNwithINchimericJJVLPsNNSmixedVBNwithINPBS (NNmeanNNtiterNN10CD\u00b1NN0).CDnsubjccconjamodamodagentpredetcasenmodaclcaseamodnmodaclcasenmodamodagentnummodamodnsubj
displaCy
Minimal or no virus-specific neutralizing antibodiesGENE_OR_GENE_PRODUCT were evident in miceORGANISM inoculated with chimeric VLPsORGANISM mixed with PBS (mean titer 10 \u00b1 0).
displaCy
TheDTmeanJJserumNNneutralizingNNtitersNNSforINtheDTgroupsNNSofINmiceNNSinoculatedVBNwithINliveJJvirus (NN70CD\u00b1NN20)CDorCCchimericJJVLPNNadministeredVBNwithINalum (NN60CD\u00b1NN23)CDwereVBDnotRBstatisticallyRBdifferentJJfromINeachDTother,JJbutCCwereVBDstatisticallyRBsignificantlyRBhigher (JJRpNN<-LRB-0.05)CDthanINthoseDTofINtheDTmiceNNSimmunizedVBNwithINaDTmixtureNNofINchimericJJVLPsNNSandCCPBSNNandCCtheDTplaceboNNgroup.NNdetamodcompoundcompoundnsubjcasedetnmodcasenmodaclcaseamodnmodnummodagentnsubjccamodconjaclcasenmodnummodagentnsubjpredetnegadvmodcasedetnmodccpredetadvmodadvmodconjcompoundagentnsubjcasenmodcasedetnmodaclcasedetnmodcaseamodnmodccconjccdetcompoundconj
displaCy
The mean serumORGANISM_SUBSTANCE neutralizing titers for the groups of miceORGANISM inoculated with live virus (70 \u00b1 20) or chimeric VLPCELL administered with alumSIMPLE_CHEMICAL (60 \u00b1 23) were not statistically different from each other, but were statistically significantly higher (p < 0.05) than those of the miceORGANISM immunized with a mixture of chimeric VLPsCELL and PBS and the placebo group.
displaCy
AfterINchallenge,NNtheDTmiceNNSimmunizedVBNwithINVeroNNE6NNcellNNcultureNNfluid (NNplacebo)NNorCCwithINaDTmixtureNNofIN2CDgNNchimericJJVLPNNandCCPBSNNhadVBDhighJJmeanNNvirusNNtitersNNSofIN6.5CDlogNN10CD/lungNNandCC5.2CDlogNN10CD/lung,NNrespectively.RBcasenmoddetnsubjaclcasecompoundcompoundcompoundcompoundnmodapposcccasedetconjcasenummodnmodamodagentccconjamodcompoundcompounddobjcasenummodcompoundnummodnmodccnummodcompoundnummodconjadvmod
displaCy
After challenge, the miceORGANISM immunized with Vero E6 cell cultureORGANISM fluid (placebo) or with a mixture of 2 g chimeric VLPCELL and PBS had high mean virus titers of 6.5 log 10 /lung and 5.2 log 10 /lung, respectively.
displaCy
InINcontrast,NNnoDTvirusNNwasVBDdetectableJJinINtheDTlungsNNSofINtheDTmiceNNSpreviouslyRBadministeredVBNeitherCCliveVBSCoVNNorCC2CDgNNofINchimericJJVLPNNmixedVBNwithINalum.NNcasenmodnegnsubjpredetcasedetnmodcasedetnmodadvmodaclnmod:npmodamoddobjccnummodconjcaseamodnmodadvmodcasenmod
displaCy
In contrast, no virus was detectable in the lungsORGAN of the miceORGANISM previously administered either live SCoVSIMPLE_CHEMICAL or 2 g of chimeric VLPCELL mixed with alumSIMPLE_CHEMICAL.
displaCy
TheDTSCoVNNtitersNNSinINtheDTlungsNNSofINtheseDTmiceNNStwoCDdaysNNSpostVBPvirusNNchallengeNNwereVBDinverselyRBproportionalJJtoTOtheirPRP$serumNNvirus-neutralizingJJantibodyNNtiters.NNSdetamodnsubjcasedetnmodcasedetnmodnummodcompoundcompoundcompoundnmodpredetadvmodcasecc:preconjcompoundamodcompoundnmod
displaCy
The SCoVSIMPLE_CHEMICAL titers in the lungsORGAN of these miceORGANISM two days post virus challenge were inversely proportional to their serumORGANISM_SUBSTANCE virus-neutralizing antibody titers.
displaCy
"''section": "NNSerumNNneutralizingNNantibodyNNtitersNNSandCClungNNSCoVNNtitersNNSinINtheDTmiceNNSimmunizedVBNonceRBwithINchimericJJVLPs"NNS_SPpunctnmodcompoundcompoundcompoundcccompoundamodconjcasedetnmodacladvmodcaseamodpunct
displaCy
"section": "SerumORGANISM_SUBSTANCE neutralizing antibody titers and lung SCoVMULTI-TISSUE_STRUCTURE titers in the miceORGANISM immunized once with chimeric VLPsCELL"
displaCy
"''ToTOstudyVBtheDTimmunogenicityNNofINchimericJJVLPs,NNSthreeCDgroupsNNSofINmiceNNSwereVBDinoculatedVBNi.m.NNpunctmarkadvcldetdobjcaseamodnmodnummodnsubjpasscasenmodauxpassdobj
displaCy
"To study the immunogenicity of chimeric VLPsCELL, three groups of miceORGANISM were inoculated i.m.
displaCy
withIN0.5,CD1,CDorCC2CDgNNmixedVBNwithINalum.NNcaseconjccnummodconjaclcasenmod
displaCy
with 0.5, 1, or 2 g mixed with alumSIMPLE_CHEMICAL.
displaCy
OtherJJgroupsNNSofINmiceNNSreceivedVBDaDTmixtureNNofIN2CDgNNofINchimericJJVLPsNNSandCCPBS,NN2CDgNNofINMHVNNVLPsNNSandCCalum,NN1CDgNNofINInfluenzaNNANNvirusNNvaccineNNandCCalum,NNVeroNNE6NNcellNNcultureNNfluid,NNliveJJSCoV,NNorCCalumNNalone,RBorCCtheDTmiceNNSreceivedVBDnoDTtreatment.NNamodnsubjcasenmoddetdobjcasenummodnmodcaseamodnmodccconjnummodnsubjcasecompoundnmodccconjnummodapposcasecompoundcompoundcompoundnmodccconjcompoundcompoundcompoundcompoundapposamodconjccconjadvmodccdetnsubjconjnegdobj
displaCy
Other groups of miceORGANISM received a mixture of 2 g of chimeric VLPsCELL and PBS, 2 g of MHV VLPsORGANISM and alumSIMPLE_CHEMICAL, 1 g of Influenza A virus vaccineORGANISM and alumSIMPLE_CHEMICAL, Vero E6 cellCELL culture fluid, live SCoVSIMPLE_CHEMICAL, or alumSIMPLE_CHEMICAL alone, or the miceORGANISM received no treatment.
displaCy
MeanNNneutralizingVBGtitersNNSatIN28CDdaysNNSpostVBPinoculationNNinINmiceNNSreceivingVBG0.5,CD1,CDorCC2CDgNNchimericJJVLPsNNSwithINalumNNwereVBD20CD\u00b1NN10,CD84CD\u00b1NN75,CDandCC110CD\u00b1NN60,CDrespectively (RBFig.NN3A) .NNamodamodnsubjcasenummodcompoundcompoundnmodcasenmodaclnummodconjccconjcompoundamoddobjcasenmodpredetnummodadvmodagentnummodamodccnummodamodnsubjadvmodcompoundconj
displaCy
Mean neutralizing titers at 28 days post inoculation in miceORGANISM receiving 0.5, 1, or 2 g chimeric VLPsCELL with alumSIMPLE_CHEMICAL were 20 \u00b1 10, 84 \u00b1 75, and 110 \u00b1 60, respectively (Fig. 3A) .
displaCy
InINtheDTabsenceNNofINalum,NN2CDgNNchimericJJVLPsNNSinducedVBDaDTlowerJJRbutCCsignificantJJneutralizingVBGantibodyNNresponseNNofIN12.5CD\u00b1NN5.CDcaseexplexplcasenmodnummodcompoundamodnsubjdetamodccconjconjcompounddobjcasenummodnmodnsubj
displaCy
In the absence of alumSIMPLE_CHEMICAL, 2 g chimeric VLPsCELL induced a lower but significant neutralizing antibody response of 12.5 \u00b1 5.
displaCy
TheDTmiceNNSinfectedVBNwithINSCoVNNhadVBDtheDTneutralizingVBGtiterNNofIN70CD\u00b1NN20.CDdetnsubjaclcasenmoddetamoddobjcasenummodnmodnsubj
displaCy
The miceORGANISM infected with SCoVSIMPLE_CHEMICAL had the neutralizing titer of 70 \u00b1 20.
displaCy
TheDTmiceNNSinoculatedVBNwithIN2CDgNNofINMHVNNVLPNNhadVBDnoDTdetectableJJvirus-specificJJneutralizingVBGantibodies,NNSdemonstratingVBGthatINMHVNNSNNproteinNNdidVBDnotRBelicitVBneutralizingVBGantibodiesNNSagainstINSCoV.NNTheDTneutralizingVBGantibodyNNtitersNNSatIN28CDdaysNNSpostVBPinoculationNNwereVBDnotRBinvestigatedVBNinINtheDTgroupsNNSofINmiceNNSinoculatedVBNwithINInfluenzaNNvaccineNNandCCalum,NNalumNNalone,RBorCCinINthoseDTreceivingVBGnoDTtreatment.NNdetnsubjaclcasenummodnmodcasecompoundnmodnegamodamodamoddobjadvclmarkcompoundcompoundnsubjauxnegccompamoddobjcasecompounddetamodcompoundnmodcasenummodcompoundcompoundnmodauxpassnegconjcasedetnmodcasenmodaclcasecompoundnmodccconjapposadvmodcccaseconjaclnegdobj
displaCy
The miceORGANISM inoculated with 2 g of MHV VLPORGANISM had no detectable virus-specific neutralizing antibodiesGENE_OR_GENE_PRODUCT, demonstrating that MHV SGENE_OR_GENE_PRODUCT protein did not elicit neutralizing antibodies against SCoV.GENE_OR_GENE_PRODUCT The neutralizing antibody titers at 28 days post inoculation were not investigated in the groups of miceORGANISM inoculated with Influenza vaccine and alumSIMPLE_CHEMICAL, alumSIMPLE_CHEMICAL alone, or in those receiving no treatment.
displaCy
"``EffectsNNSofINchimericJJVLPNNamountsNNSonINneutralizingVBGantibodyNNtitersNNSandCCinhibitionNNofINchallengedVBNSCoVNNreplicationNNinINtheDTlungs"NNS_SPpunctcaseamodcompoundnmodcaseamodcompoundnmodccconjcaseamodamodnmodcasedetpunct
displaCy
"Effects of chimeric VLPCELL amounts on neutralizing antibody titers and inhibition of challenged SCoVSIMPLE_CHEMICAL replication in the lungsORGAN"
displaCy
AtINdayNN28,CDtheseDTmice,NNSexceptINforINthoseDTuntreatedJJandCCthoseDTthatWDThadVBDbeenVBNinoculatedVBNwithINliveJJSCoV,NNwereVBDboostedVBNwithINtheDTsameJJmaterial.NNpreppobjnsubjdetnsubjpasscasecasenmodaclccconjnsubjpassauxauxpassparataxisprepamodpobjauxpasscasedetamodnmod
displaCy
At day 28, these miceORGANISM, except for those untreated and those that had been inoculated with live SCoVSIMPLE_CHEMICAL, were boosted with the same material.
displaCy
MiceNNSinjectedVBNwithINchimericJJVLPsNNSandCCalumNNhadVBDnoDTincreaseNNinINtiterNNatINtheDTlowestJJSdose,NNbutCC1-andJJ2-gJJdosesNNSresultedVBDinINmarkedJJboosterNNresponses.NNSnsubjaclcaseamodnmodccconjnegdobjcasenmodcasedetamodnmodccamodamodnsubjconjcaseamodcompoundnmod
displaCy
MiceORGANISM injected with chimeric VLPs and alumSIMPLE_CHEMICAL had no increase in titer at the lowest dose, but 1-and 2-g doses resulted in marked booster responses.
displaCy
AnimalsNNSreceivingVBGtheDThighestJJSdoseNNdevelopedJJtitersNNSofIN200CD\u00b1NN97.7,CDwhileINthoseDTinfectedVBNwithINSCoVNNhadVBDtheDTneutralizingVBGtiterNNofIN97CD\u00b1NN60.CDnsubjacldetamoddobjdobjcasenummodamodnmodmarknsubjaclcasenmodadvcldetamoddobjcasenummodnmodnsubj
displaCy
Animals receiving the highest dose developed titers of 200 \u00b1SIMPLE_CHEMICAL 97.7, while those infected with SCoVSIMPLE_CHEMICAL had the neutralizing titer of 97 \u00b1 60.
displaCy
ADTsecondJJinoculationNNofINaDTmixtureNNofIN2CDgNNchimericJJVLPsNNSwithINPBSNNefficientlyRBboostedVBDmedianJJneutralizingVBGantibodyNNtitersNNStoTO57CD\u00b1NN21,CDwhichWDTwasVBDhigherJJRthanINtheDTtiterNNfollowingVBG0.5CDgNNofINchimericJJVLPNNmixedVBNwithINalum (NN15CD\u00b1NN5.7).CDdetamodnsubjcasedetnmodcasenummodcompoundamodnmodcasenmodadvmodamodamodcompounddobjcasenummodamodnmodnsubjpredetparataxiscasedetnmodcasenummodnmodcaseamodnmodaclcasenmodnummodagentnsubj
displaCy
A second inoculation of a mixture of 2 g chimeric VLPs with PBS efficiently boosted median neutralizing antibody titers to 57 \u00b1 21, which was higher than the titer following 0.5 g of chimeric VLPCELL mixed with alumSIMPLE_CHEMICAL (15 \u00b1 5.7).
displaCy
NoDTneutralizingVBGantibodiesNNSwereVBDdetectedVBNatINtheDTlowestJJSdilutionNNtestedVBNinINgroupsNNSreceivingVBGMHVNNVLP,NNinfluenzaNNvaccineNNandCCalum,NNalumNNalone,RBorCCplaceboNNorCCinINthoseDTleftJJuntreated.JJnegamodnsubjpassauxpasscasedetamodnmodaclcasenmodaclcompounddobjcompoundconjccconjapposadvmodccconjcccaseconjaclxcomp
displaCy
No neutralizing antibodies were detected at the lowest dilution tested in groups receiving MHV VLPORGANISM, influenza vaccine and alumSIMPLE_CHEMICAL, alumSIMPLE_CHEMICAL alone, or placebo or in those leftORGAN untreated.
displaCy
"``EffectsNNSofINchimericJJVLPNNamountsNNSonINneutralizingVBGantibodyNNtitersNNSandCCinhibitionNNofINchallengedVBNSCoVNNreplicationNNinINtheDTlungs"NNS_SPpunctcaseamodcompoundnmodcaseamodcompoundnmodccconjcaseamodamodnmodcasedetpunct
displaCy
"Effects of chimeric VLPCELL amounts on neutralizing antibody titers and inhibition of challenged SCoVSIMPLE_CHEMICAL replication in the lungsORGAN"
displaCy
"``AllDTmiceNNSwereVBDchallengedVBNi.n.NNSpunctdetnsubjpassauxpassdobj
displaCy
"All miceORGANISM were challenged i.n.
displaCy
withINliveJJSCoVNNonINdayNN56CDandCCsacrificedVBD2CDdaysNNSlater.RBcaseamodcasenmodnsubjccconjnummodnpadvmodadvmod
displaCy
with live SCoVSIMPLE_CHEMICAL on day 56 and sacrificed 2 days later.
displaCy
AsINexpected,VBNmaximalJJpulmonaryJJvirusNNtitersNNSwereVBDdetectedVBNinINtheDTgroupsNNSofINmiceNNSinoculatedVBNwithINplacebo (NN6.9CDlogNN10CD\u00b10.7),NNMHVNNVLPs (NNS6.2CDlogNN10CD\u00b10.4),NNinfluenzaNNvaccineNNandCCalum (NN7.5CD\u00b1NN0.1),CDalumNNalone (RB7.3CD\u00b1NN0),CDandCCinINthoseDTleftJJuntreated (JJ8.5CD\u00b1NN0) (CDFig.NN3C) .NNmarkadvclamodamodcompoundnsubjpassauxpasscasedetnmodcasenmodaclcasenmodnummodagentnummodagentcompoundconjnummodagentnsubjnsubjcompoundconjccconjnummodamodapposapposadvmodnummodagentnsubjcccaseconjamodamodnummodexplnsubjcompoundappos
displaCy
As expected, maximal pulmonary virusCELL titers were detected in the groups of miceORGANISM inoculated with placebo (6.9 log 10 \u00b10.7), MHV VLPsORGANISM (6.2 log 10 \u00b10.4GENE_OR_GENE_PRODUCT), influenza vaccine and alumSIMPLE_CHEMICAL (7.5 \u00b1 0.1), alumSIMPLE_CHEMICAL alone (7.3 \u00b1 0), and in those leftORGAN untreated (8.5 \u00b1 0) (Fig. 3C) .
displaCy
PartialJJprotectionNNwasVBDobtainedVBNinINgroupsNNSofINmiceNNSinoculatedVBNtwiceRBwithIN0.5CDgNNchimericJJVLPNNmixedVBNwithINalum (NN5.5CDlogNN10CD\u00b10.6),NN2CDgNNchimericJJVLPsNNSandCCPBS (NN3.9CDlogNN10CD\u00b11.1)NNorCC1CDgNNchimericJJVLPsNNSmixedVBNwithINalum (NN3.1CDlogNN10CD\u00b10.5).NNSamodnsubjpassauxpasscasenmodcasenmodacladvmodcasenummodnmodamodagentadvmodcasenmodnummodcompoundnummodagentnummodnmodamodagentccconjnummodexplnummodapposccnummodconjamodagentaclcasenmodnummodagentnummodnsubj
displaCy
Partial protection was obtained in groups of miceORGANISM inoculated twice with 0.5 g chimeric VLPCELL mixed with alumSIMPLE_CHEMICAL (5.5 log 10 \u00b10.6GENE_OR_GENE_PRODUCT), 2 g chimeric VLPsCELL and PBS (3.9 log 10 \u00b11.1) or 1 g chimeric VLPsORGANISM mixed with alumSIMPLE_CHEMICAL (3.1 log 10 \u00b10.5GENE_OR_GENE_PRODUCT).
displaCy
Notably,RBnoDTvirusNNSCoV-specificJJneutralizingVBGserumNNantibodyNNtitersNNSinINtheDTimmunizedJJmiceNNSandCCSCoVNNtitersNNSinINlungsNNSofINmiceNNS2CDdaysNNSafterINintranasalJJchallengeNNwithINSCoVNNatINdayNN56.CDadvmodnegcompoundamodamodcompoundcompoundcasedetamodnmodccamodconjcasenmodcasenmodnummodconjcaseamodnmodcasenmodcasenmodnsubj
displaCy
Notably, no virus SCoV-specific neutralizing serumORGANISM_SUBSTANCE antibody titers in the immunized miceORGANISM and SCoVSIMPLE_CHEMICAL titers in lungsORGAN of mice 2ORGANISM days after intranasal challenge with SCoVSIMPLE_CHEMICAL at day 56.
displaCy
OnINdayNN0,CDmiceNNSwereVBDeitherRBleftVBNuntreatedJJorCCwereVBDinoculatedVBNi.n.NNSpreppobjnsubjnsubjpassauxpassnmod:npmodxcompccauxpassconjdobj
displaCy
On day 0, miceORGANISM were either leftORGAN untreated or were inoculated i.nORGANISM.
displaCy
withIN1CD\u00d7NN10CD6CDTCIDNN50CDofINSCoVNNorCCinjectedJJi.m.NNcasenummodexplnummodnummodnsubjcasenmodccconjagent
displaCy
with 1 \u00d7 10 6 TCID 50 of SCoVSIMPLE_CHEMICAL or injected i.mSIMPLE_CHEMICAL.
displaCy
withINplacebo (NNmediumNNfromINVeroNNE6NNcells),NNSaDTmixtureNNofIN2CDgNNMHVNNVLPsNNSandCCPBS,NNaDTmixtureNNofIN2CDgNNchimericJJVLPNNandCCPBS,NNaDTmixtureNNofIN2CDgNNchimericJJVLPNNandCCalum,NNaDTmixtureNNofIN1CDgNNchimericJJVLPNNandCCalum,NNaDTmixtureNNofIN0.5CDgNNchimericJJVLPNNandCCalum,NNaDTmixtureNNofIN1CDgNNinfluenzaNNvirusNNvaccineNNandCCalum,NNorCCalumNNalone.RBcaseagentcasecompoundcompoundnmoddetapposcasenummodnmodcompoundagentccconjdetapposcasenummodnmodamodagentccconjdetapposcasenummodnmodamodagentccconjdetapposcasenummodnmodamodagentccconjdetapposcasenummodnmodamodagentccconjdetapposcasenummodcompoundcompoundcompoundnmodccconjccconjadvmod
displaCy
with placebo (medium from Vero E6 cellsCELL), a mixture of 2 g MHV VLPsORGANISM and PBS, a mixture of 2 g chimeric VLPCELL and PBS, a mixture of 2 g chimeric VLPCELL and alumSIMPLE_CHEMICAL, a mixture of 1 g chimeric VLPCELL and alumSIMPLE_CHEMICAL, a mixture of 0.5 g chimeric VLPCELL and alumSIMPLE_CHEMICAL, a mixture of 1 g influenza virus vaccine and alumSIMPLE_CHEMICAL, or alumSIMPLE_CHEMICAL alone.
displaCy
(-LRB-A)LSTheDTgraphNNrepresentsVBZvirus-specificJJserumNNneutralizingNNantibodyNNtitersNNSatIN28CDdaysNNSpostVBPinoculation.NNpunctexpldetnsubjamodcompoundcompoundcompounddobjcasenummodcompoundcompoundnmod
displaCy
(A) The graph represents virus-specific serumORGANISM neutralizing antibody titers at 28 days post inoculation.
displaCy
(-LRB-B)NNAtIN56CDdaysNNSbloodNNsamplesNNSwereVBDcollected,VBNandCCthenRBtheDTmice,NNSexcludingVBGthoseDTinoculatedVBNwithINliveJJSCoVNNandCCleftVBDuntreated,JJwereVBDre-inoculatedVBNwithINtheDTsameJJmaterial.NNpunctexplcasecompoundnmodcompoundnsubjpassauxpassccadvmoddetnsubjpasscasenmodaclcaseamodnmodccconjxcompauxpassconjcasedetamodnmod
displaCy
(B) At 56 days blood samplesORGANISM_SUBSTANCE were collected, and then the miceORGANISM, excluding those inoculated with live SCoVSIMPLE_CHEMICAL and leftORGAN untreated, were re-inoculated with the same material.
displaCy
TheDTgraphNNrepresentsVBZvirus-specific,JJserumneutralizingVBGantibodyNNtitersNNSatIN56CDdaysNNSafterINinitialJJinoculation.NNdetnsubjamodamodcompounddobjcasenummodnmodcaseamodnmod
displaCy
The graph represents virus-specificGENE_OR_GENE_PRODUCT, serumneutralizing antibody titers at 56 days after initial inoculation.
displaCy
(-LRB-C)NNMiceNNSwereVBDinoculatedVBNwithIN1CD\u00d7NN10CD6CDTCIDNN50CDofINSCoVNNatINdayNN56.CDpunctcompoundnsubjpassauxpasscasenummodexplnummodnummodnmodnsubjcasenmodcasenmodnsubj
displaCy
(C) Mice were inoculated with 1 \u00d7 10 6 TCID 50 of SCoVSIMPLE_CHEMICAL at day 56.
displaCy
AfterIN2CDdays,NNSmiceNNSwereVBDeuthanized,VBNandCCtheDTlungNNvirusNNtitersNNSwereVBDdetermined.VBNcasenummodnmodnsubjpassauxpassccdetcompoundcompoundnsubjpassauxpassconj
displaCy
After 2 days, miceORGANISM were euthanized, and the lung virusTISSUE titers were determined.
displaCy
TheDTverticalJJdashedVBDlineNNin (INC)NNdenotesVBZtheDTminimalJJvirusNNdetectionNNlevelNNinINthisDTassay (NN2.3CDlogNN10CDTCIDNN50CD/gNNlung).NNdetamodamodnsubjcasenmoddetamodcompoundcompounddobjcasedetnmodnummodcompoundnummodcompoundcompoundcompoundappos
displaCy
The vertical dashed line in (C) denotes the minimal virus detection level in this assay (2.3 log 10 TCID 50 /g lungORGAN).
displaCy
TheDTnumberNNofINmiceNNSusedVBNfor (INB)NNand (CCC)NNwere:VBD7CDforINplacebo,NNSCoV,NNandCC2CDgNNchimericJJVLPNNwithINPBS;NN6CDforIN2CDgNNchimericJJVLPNNwithINalum;NN5CDforINMHVNNVLPNNwithINalumNNandCC1CDgNNchimericJJVLPNNwithINalum;NN4CDforIN0.5CDgNNchimericJJVLPNNwithINalum;NNandCC3CDforINallDTotherJJgroups.NNSdetnsubjcasenmodaclcasenmodccappospredetcasenmodconjccnummodconjamodagentcasenmodagentcasenummodnmodamodnmodcasenmodagentcasecompoundnmodcasenmodccnummodconjamodnmodcasenmodnummodcasenummodnmodamodagentcasenmodccconjcasedetamodnmod
displaCy
The number of miceORGANISM used for (B) and (C) were: 7 for placebo, SCoVSIMPLE_CHEMICAL, and 2 g chimeric VLPCELL with PBS; 6 for 2 g chimeric VLPCELL with alumSIMPLE_CHEMICAL; 5 for MHV VLPORGANISM with alumSIMPLE_CHEMICAL and 1 g chimeric VLPCELL with alumSIMPLE_CHEMICAL; 4 for 0.5 g chimeric VLPCELL with alumSIMPLE_CHEMICAL; and 3 for all other groups.
displaCy
wasVBDdetectableJJinINtheDTlungsNNSofINanyDTofINtheDTmiceNNSgivenVBNliveJJvirusNNi.n.NNSpredetcasedetnmodcasenmodcasedetnmodaclamodcompounddobj
displaCy
was detectable in the lungsORGAN of any of the miceORGANISM given live virus i.nORGANISM.
displaCy
orCCtwoCDdosesNNSofIN2CDgNNchimericJJVLPNNmixedVBNwithINalum.NNccnummodcasenummodnmodamodagentnummodcasenmod
displaCy
or two doses of 2 g chimeric VLPCELL mixed with alumSIMPLE_CHEMICAL.
displaCy
TheDTmeanJJlungNNvirusNNtitersNNSforINtheDTlatterJJtwoCDgroupsNNSwereVBDstatisticallyRBdifferentJJfromINtheDTmeanJJpulmonaryJJSCoVNNtiterNNseenVBNinINtheDTplaceboNNcontrolNNgroupNNwhenWRBtheseDTmeansNNSwereVBDcomparedVBNusingVBGaDTnon-parametricJJANOVANNtest (NNbothDTpNNvaluesNNSbeingVBG<NN0.01).CDdetamodcompoundcompoundnsubjcasedetamodnummodnmodpredetadvmodcasedetamodamodamodnmodaclcasedetcompoundcompoundnmodadvmoddetnsubjpassauxpassadvclxcompdetamodcompounddobjdetcompoundnsubjpredetcompoundappos
displaCy
The mean lung virus titers for the latter two groups were statistically different from the mean pulmonary SCoVMULTI-TISSUE_STRUCTURE titer seen in the placebo control group when these means were compared using a non-parametric ANOVA test (both p values being <0.01).
displaCy
Overall,RBtheDTvirusNNlevelsNNSinINtheseDTanimalsNNSgenerallyRBcorrelatedVBDinverselyRBwithINthoseDTofINSCoVspecificJJneutralizingVBGantibodiesNNSpresentJJinINtheDTseraNNofINtheDTmice.NNSadvmoddetcompoundnsubjcasedetnmodadvmodadvmodcasenmodcaseamodamodnmodnummodcasedetnmodcasedetnmod
displaCy
Overall, the virus levels in these animals generally correlated inversely with those of SCoVspecific neutralizing antibodies present in the seraORGANISM_SUBSTANCE of the miceORGANISM.
displaCy
"``EffectsNNSofINchimericJJVLPNNamountsNNSonINneutralizingVBGantibodyNNtitersNNSandCCinhibitionNNofINchallengedVBNSCoVNNreplicationNNinINtheDTlungs"NNS_SPpunctcaseamodcompoundnmodcaseamodcompoundnmodccconjcaseamodamodnmodcasedetpunct
displaCy
"Effects of chimeric VLPCELL amounts on neutralizing antibody titers and inhibition of challenged SCoVSIMPLE_CHEMICAL replication in the lungsORGAN"
displaCy
"``HistopathologicalJJexaminationNNofINtheDTgroupsNNSofINmiceNNSthatWDTneitherCCproducedVBDneutralizingVBGantibodiesNNSnorCCsuppressedVBDSCoVNNreplicationNNinINtheDTlungs (NNSi.e.,FWmiceNNStreatedVBNwithINVeroNNE6NNcellNNcultureNNfluid,NNinoculatedVBNwithINaDTmixtureNNofINinfluenzaNNvirusNNvaccineNNandCCalum,NNandCCwithINalumNNalone,RBasRBwellRBasINthoseDTleftJJuntreated),JJrevealedVBDmoderateJJinterstitialJJpneumonia (NNFig.NN4ANNandCCB) .NNpunctamodnsubjcasedetnmodcasenmodmarknmod:npmodamodamodnsubjccparataxisamoddobjcasedetnmodexplnmodaclcasecompoundcompoundcompoundcompoundnmodaclcasedetnmodcasecompoundcompoundnmodccconjcccaseconjadvmodccquantmodquantmodconjaclxcompamodamoddobjcompoundagentccconj
displaCy
"Histopathological examination of the groups of miceORGANISM that neither produced neutralizing antibodies nor suppressed SCoVSIMPLE_CHEMICAL replication in the lungsORGAN (i.e., miceORGANISM treated with Vero E6 cellCELL culture fluid, inoculated with a mixture of influenza virus vaccine and alumSIMPLE_CHEMICAL, and with alumSIMPLE_CHEMICAL alone, as well as those leftORGAN untreated), revealed moderate interstitial pneumonia ( Fig. 4A and BCELL) .
displaCy
TheDTbronchialJJepitheliumNNappearedVBDtoTObeVBtheDTmainJJaffectedVBNtargetNNwithINprominentJJcellularJJcytoplasmicJJswellingNNandCCblebbing.NNdetamodnsubjmarkpredetdetamodamodxcompcaseamodamodamodnmodccconj
displaCy
The bronchial epitheliumTISSUE appeared to be the main affected target with prominent cellular cytoplasmicTISSUE swelling and blebbing.
displaCy
ExtensiveJJaccumulationNNofINcellularJJdebrisNNandCCnecroticJJepithelialJJcells,NNSaccompaniedVBNbyINinflammatoryJJinfiltrates,NNSoccurredVBDinINsomeDTbronchioles (NNSFig.NN4A) .NNamodnsubjcaseamodnmodccamodamodconjaclcaseamodnmodcasedetnmodcompoundappos
displaCy
Extensive accumulation of cellularCELL debris and necrotic epithelial cellsCELL, accompanied by inflammatory infiltrates, occurred in some bronchiolesTISSUE (Fig. 4A) .
displaCy
WePRPalsoRBobservedVBDaDTmoderateJJinfiltrationNNofINmononuclearJJcellsNNSaroundINperibronchiolarJJandCCperivascularJJregionsNNSofINinfectedJJtissuesNNSandCCaDTmild-to-moderateJJthickeningNNofINtheDTbronchiolarJJinterstitialJJtissuesNNSandCCalveolarJJwallsNNSwithINmononuclearJJcellNNinfiltration (NNFig.NN4B) .NNnsubjadvmoddetamoddobjcaseamodnmodcasenmodccamodconjcaseamodnmodccdetamodconjcasedetamodamodnmodccamodconjcaseamodcompoundnmodcompoundappos
displaCy
We also observed a moderate infiltration of mononuclear cellsCELL around peribronchiolarTISSUE and perivascular regionsTISSUE of infected tissuesTISSUE and a mild-to-moderate thickening of the bronchiolar interstitial tissuesTISSUE and alveolar wallsTISSUE with mononuclear cellCELL infiltration (Fig. 4B) .
displaCy
IHCNNstainingNNdemonstratedVBDtheDTpresenceNNofINSCoVNNNNNproteinNNwithinINbronchiolarJJepithelialJJcells,NNSbutCCnotRBwithinINcellsNNSofINalveolarJJlining (NNFig.NN4C) .NNcompoundnsubjdetdobjcaseamodcompoundnmodcaseamodamodnmodccnegcaseconjcaseamodnmodcompoundappos
displaCy
IHC staining demonstrated the presence of SCoV NGENE_OR_GENE_PRODUCT protein within bronchiolar epithelial cellsCELL, but not within cellsCELL of alveolar liningMULTI-TISSUE_STRUCTURE (Fig. 4C) .
displaCy
InINcontrastNNtoTOtheDTprominentJJpathologyNNandCCviralJJreplicationNNinINtheDTlungsNNSofINtheDTcontrolJJanimals,NNSmildJJinterstitialJJpneumoniaNNwasVBDfound,VBNuponINviralJJchallenge,NNinINtheDTmiceNNSpreviouslyRBimmunizedVBNwithINmixturesNNSofINeitherCCchimericJJVLPNNandCCPBS,NNorCCchimericJJVLPNNandCCalum,NNandCCinINthoseDTinfectedVBNwithINliveJJSCoV (NNFig.NN4D,NNE,NNG,NNH) .NNcaseexplcasedetamodnmodccamodconjcasedetnmodcasedetcompoundnmodamodamodnsubjpassauxpasscaseamodnmodcasedetnmodadvmodaclcasenmodcasenmod:npmodamodnmodccconjccamodconjccconjcccaseconjaclcaseamodnmodcompoundagentapposapposappos
displaCy
In contrast to the prominent pathology and viral replication in the lungsORGAN of the control animals, mild interstitial pneumonia was found, upon viral challenge, in the miceORGANISM previously immunized with mixtures of either chimeric VLPCELL and PBS, or chimeric VLPCELL and alumSIMPLE_CHEMICAL, and in those infected with live SCoVSIMPLE_CHEMICAL (Fig. 4D, E, G, H) .
displaCy
Specifically,RBcytopathology,NNi.e.,FWswellingNNandCCblebbing,NNwasVBDrarelyRBobservedVBNinINbronchiolarJJepithelialJJcells,NNSdespiteINtheDTpresenceNNofINsomeDTdesquamatedJJcellsNNSwithinINtheDTairwayNNlumen.NNadvmodnsubjpassadvmodagentccconjauxpassadvmodcaseamodamodnmodcaseexplexplcasedetamodnmodcasedetcompoundnmod
displaCy
Specifically, cytopathologyPATHOLOGICAL_FORMATION, i.e., swelling and blebbingCELLULAR_COMPONENT, was rarely observed in bronchiolar epithelial cellsCELL, despite the presence of some desquamated cellsCELL within the airway lumenMULTI-TISSUE_STRUCTURE.
displaCy
Additionally,RBtheDTthickeningNNofINbronchiolarJJinterstitialJJtissuesNNSandCCtheDTalveolarJJwallNNandCCcellularJJinfiltrationNNwereVBDlessRBRprominentJJwhenWRBcomparedVBNwithINthoseDTofINtheDTcontrolJJgroups.NNSadvmoddetnsubjcaseamodamodnmodccdetamodconjccamodconjpredetadvmodadvmodadvclcasenmodcasedetcompoundnmod
displaCy
Additionally, the thickening of bronchiolar interstitial tissuesTISSUE and the alveolar wallTISSUE and cellularCELL infiltration were less prominent when compared with those of the control groups.
displaCy
ThereEXwasVBDnoDTbigJJdifferenceNNinINtheDTlungNNpathologyNNbetweenINmiceNNSinoculatedVBNwithINVLPNNandCCPBSNNandCCthoseDTinoculatedVBNwithINVLPNNandCCalum.NNdobjnegamodnegcasedetcompoundnmodcasenmodaclcasenmodccconjccconjaclcasenmodccconj
displaCy
There was no big difference in the lungORGAN pathology between miceORGANISM inoculated with VLPCELL and PBS and those inoculated with VLPSIMPLE_CHEMICAL and alumSIMPLE_CHEMICAL.
displaCy
WePRPnoticedVBDthatINinflammatoryJJmononuclearJJcellsNNSseemedVBDtoTObeVBtheDTmain,JJifINnotRBonly,RBcellularJJcomponentNNofINtheDTcellularJJinfiltratesNNSaroundINtheDTperibronchiolarJJandCCperivascularJJregionsNNSofINmiceNNSinitiallyRBprimedVBNwithINliveJJSCoV.",NN_SPnsubjmarkamodamodnsubjccompmarkpredetdetamodmarknegagentamodnmodcasedetamodnmodcaseexplnmodccamodconjcasenmodadvmodaclcaseamodpunct
displaCy
We noticed that inflammatory mononuclear cellsCELL seemed to be the main, if not only, cellularCELL component of the cellular infiltratesTISSUE around the peribronchiolarTISSUE and perivascular regionsTISSUE of miceORGANISM initially primed with live SCoV.GENE_OR_GENE_PRODUCT",
displaCy
"''section": "NNHistopathologicalJJexaminationNNandCCIHCNNofINtheDTlungsNNSofINtheDTimmunizedJJmiceNNSafterINSCoVNNchallenge"NN_SPpunctamodagentccconjcasedetnmodcasedetamodnmodcaseamodpunct
displaCy
"section": "Histopathological examination and IHC of the lungsORGAN of the immunized miceORGANISM after SCoVSIMPLE_CHEMICAL challenge"
displaCy
InINadditionNNtoTOinflammatoryJJmononuclearJJcells,NNSwePRPobservedVBDtheDTinfiltrationNNofINneutrophilsNNSandCCeosinophilsNNSaroundINtheDTbronchiolesNNSandCCbloodNNvesselsNNSofINmiceNNSinoculatedVBNwithINaDTmixtureNNofINchimericJJVLPsNNSandCCPBSNNandCCthatDTofINchimericJJVLPsNNSandCCalum (NNFig.NN4DNNandCCE) ;NNcountingNNofINtheDTinfiltratingVBGcellsNNSatINtheDTfiveCDfieldsNNSofINeachDTmouseNN'sPOSlungNNtissuesNNSrevealedVBDthatINinfiltratingVBGeosinophilsNNSrepresentedVBD13.2CD\u00b1NN9.6%CDandCC22.2CD\u00b1NN9.9%CDofINallPDTtheDTinfiltratingVBGcellsNNSinINmiceNNSinoculatedVBNwithINaDTmixtureNNofINchimericJJVLPNNandCCPBSNNandCCinINthoseDTinoculatedVBNwithINchimericJJVLPNNandCCalum,NNrespectively.RBcaseexplcaseamodamodnmodnsubjdetdobjcasenmodccconjcasedetnmodcccompoundconjcasenmodaclcasedetnmodcaseamodnmodccconjccconjcaseamodnmodccconjcompoundagentccconjnsubjcasedetamodnmodcasedetnummodnmodcasedetcc:preconjcasecompoundnmodmwemarkamodnsubjccompnummodamoddobjccnummodamodconjcaseadvmoddetamodnmodcasenmodaclcasedetnmodcaseamodnmodccconjcccaseconjaclcaseamodnmodccconjadvmod
displaCy
In addition to inflammatory mononuclear cellsCELL, we observed the infiltration of neutrophilsCELL and eosinophilsCELL around the bronchiolesTISSUE and blood vesselsMULTI-TISSUE_STRUCTURE of miceORGANISM inoculated with a mixture of chimeric VLPsCELL and PBS and that of chimeric VLPsCELL and alumSIMPLE_CHEMICAL ( Fig. 4DCANCER and E) ; counting of the infiltrating cellsCELL at the five fields of each mouseORGANISM's lung tissuesTISSUE revealed that infiltrating eosinophilsCELL represented 13.2 \u00b1 9.6% and 22.2 \u00b1 9.9% of all the infiltrating cellsCELL in miceORGANISM inoculated with a mixture of chimeric VLPCELL and PBS and in those inoculated with chimeric VLPCELL and alumSIMPLE_CHEMICAL, respectively.
displaCy
InINcontrast,NNinfiltratingVBGeosinophilsNNSrepresentedVBDmerelyRB1.42CD\u00b1NN1.42%,CD0%,CD0%CDandCC0.77CD\u00b1NN0.33%CDofINallDTinfiltratingVBGcellsNNSinINmiceNNSinoculated,VBNrespectively,RBwithINVeroNNE6NNcellNNcultureNNfluid,NNaDTmixtureNNofINinfluenzaNNvirusNNvaccineNNandCCalum,NNalumNNalone,RBandCCinINthoseDTleftJJuntreated.JJcasenmodamodnsubjadvmodnmodagentagentapposapposccnummodadvmodconjcasedetamodnmodcasenmodacladvmodcasecompoundcompoundcompoundcompoundnmoddetapposcasecompoundcompoundnmodccconjapposadvmodcccaseconjaclxcomp
displaCy
In contrast, infiltrating eosinophilsCELL represented merely 1.42 \u00b1 1.42%, 0%, 0% and 0.77 \u00b1 0.33% of all infiltrating cellsCELL in miceORGANISM inoculated, respectively, with Vero E6 cellCELL culture fluid, a mixture of influenza virus vaccine and alumSIMPLE_CHEMICAL, alumSIMPLE_CHEMICAL alone, and in those leftORGAN untreated.
displaCy
"''section": "NNHistopathologicalJJexaminationNNandCCIHCNNofINtheDTlungsNNSofINtheDTimmunizedJJmiceNNSafterINSCoVNNchallenge"NN_SPpunctamodagentccconjcasedetnmodcasedetamodnmodcaseamodpunct
displaCy
"section": "Histopathological examination and IHC of the lungsORGAN of the immunized miceORGANISM after SCoVSIMPLE_CHEMICAL challenge"
displaCy
"``TheDTpresentJJstudyNNtestedVBDtheDTefficacyNNofINimmunizationNNofINmiceNNSwithINchimericJJVLPsNNSforINelicitationNNofINanti-SCoVneutralizingJJantibodies,NNSsuppressionNNofINSCoVNNreplicationNNinINtheDTlungs,NNSandCCSCoV-mediatedJJlungNNcytopathology.NNpunctdetamodnsubjdetdobjcasenmodcasenmodcaseamodnmodcasenmodcaseamodnmoddobjcaseamodnmodcasedetnmodccamodcompoundconj
displaCy
"The present study tested the efficacy of immunization of miceORGANISM with chimeric VLPs for elicitation of anti-SCoVneutralizing antibodies, suppression of SCoVSIMPLE_CHEMICAL replication in the lungsORGAN, and SCoV-mediated lung cytopathologyCANCER.
displaCy
AlthoughINvariousJJstrategiesNNSforINSCoVNNvaccines,NNSincludingVBGexpressionNNofINSCoVNNSNNproteinNNinINotherJJviruses [NNS18,CD48,CD49] ,CDinactivatedVBNSCoVNNparticles ,NNSDNANNvaccines [NNS17,CD54] ,CDrecombinantJJSNNprotein [NN55,CD56]CDandCCotherJJapproaches [NNS57,CD58] ,CDhaveVBPbeenVBNreported,VBNtoTOourPRP$knowledgeNNthisDTisVBZtheDTfirstJJreportNNtestingVBGVLPsNNSpropagatedVBNinINmammalianJJcellsNNSasINSCoVNNvaccineNNcandidates.NNSmarkamodnsubjcaseamodnmodcasenmodcaseamodcompoundnmodcaseamodnmodagentconjconjadvclamoddobjcompoundconjnummodconjamodcompoundagentnummodnsubjccamodconjagentapposauxauxpasscasecc:preconjnmodnsubjpredetdetamodconjacldobjaclcaseamodnmodcaseamodcompoundnmod
displaCy
Although various strategies for SCoV vaccinesSIMPLE_CHEMICAL, including expression of SCoV SGENE_OR_GENE_PRODUCT protein in other viruses [18, 48, 49] , inactivated SCoV particlesSIMPLE_CHEMICAL , DNACELLULAR_COMPONENT vaccines [17, 54] , recombinant S protein [55, 56] and other approaches [57, 58] , have been reported, to our knowledge this is the first report testing VLPsCELL propagated in mammalian cellsCELL as SCoV vaccineGENE_OR_GENE_PRODUCT candidates.
displaCy
ItPRPappearsVBZthatINchimericJJVLPsNNSproducedVBNfromINmammalianJJcellsNNSinINthisDTstudyNNwereVBDmoreRBRimmunogenicJJthanINwereVBDSCoVNNVLPsNNSproducedVBNbyINusingVBGaDTbaculovirus-expressionNNsystem ;NNimmunizationNNofINtheDTmiceNNSfourCDtimesNNSwithIN100CDgNNofINtheDTbaculovirus-derivedJJSCoVNNVLPsNNSinINeachDTimmunizationNNelicitedVBDneutralizingVBGantibodyNNtitersNNSthatWDTwereVBDnoRBlongerRBRhigherJJRthanINthoseDTobtainedVBNinINtheDTpresentJJstudy.NNnsubjmarkamodnsubjaclcaseamodnmodcasedetnmodpredetadvmodccompmarkpredetamodnmodaclmarkadvcldetamoddobjnsubjcasedetnmodnummodadvmodcasenummodnmodcasedetamodamodnmodcasedetnmodccompamodcompounddobjnsubjpredetnegadvmodparataxiscasenmodaclcasedetamodnmod
displaCy
It appears that chimeric VLPsCELL produced from mammalian cellsCELL in this study were more immunogenic than were SCoV VLPsORGANISM produced by using a baculovirus-expression system ; immunization of the miceORGANISM four times with 100 g of the baculovirus-derived SCoV VLPsORGANISM in each immunization elicited neutralizing antibody titers that were no longer higher than those obtained in the present study.
displaCy
UseNNofINinactivatedVBNSCoVNNasINaDTSCoVNNvaccineNNisVBZaDTsimilarJJvaccineNNstrategyNNthatWDTwasVBDexploredVBNinINtheDTpresentJJstudy.NNnsubjcaseamodnmodcasedetamodnmodpredetdetamodcompoundnsubjpassauxpassparataxiscasedetamodnmod
displaCy
Use of inactivated SCoVSIMPLE_CHEMICAL as a SCoV vaccineSIMPLE_CHEMICAL is a similar vaccine strategy that was explored in the present study.
displaCy
OthersNNSreportedVBDthatINimmunizingJJmiceNNStwiceRBwithIN1CDgNNofINdouble-inactivatedJJSCoVNNwithINalumNNelicitedVBDhighJJtitersNNSofINneutralizingVBGantibodies .NNSnsubjmarkamodnsubjadvmodcasenummodnmodcaseamodnmodcasenmodccompamoddobjcaseamodnmod
displaCy
Others reported that immunizing miceORGANISM twice with 1 g of double-inactivated SCoVSIMPLE_CHEMICAL with alumSIMPLE_CHEMICAL elicited high titers of neutralizing antibodies .
displaCy
ADTmajorJJnegativeJJaspectNNofINtheDTuseNNofINinactivatedVBNSCoVNNvaccinesNNSisVBZthatINextremeJJcareNNmustMDbeVBtakenVBNtoTOcompletelyRBinactivateVBinfectiousJJSCoVNNinINtheDTvaccineNNpreparations.NNSdetamodamodnsubjcasedetnmodcaseamodamodnmodmarkamodnsubjpassauxauxpassccompmarkadvmodxcompamoddobjcasedetcompoundnmod
displaCy
A major negative aspect of the use of inactivated SCoV vaccines is that extreme care must be taken to completely inactivate infectious SCoVSIMPLE_CHEMICAL in the vaccine preparations.
displaCy
"``AccumulatedJJdataNNSsuggestVBPtheDTimportanceNNofINtheDTcompatibilityNNofINtheDTSNNproteinNNendodomain (NNorCCaDTcytoplasmicJJtail)NNandCCMNNproteinNNforINassemblyNNofINSNNproteinNNintoINcoronavirusNNorCCVLPs [NNS40,CD59,CD60] ;CDinINpastJJreports,NNSSNNproteinNNassemblyNNoccurredVBDifINtheseDTtwoCDregionsNNSareVBPderivedVBNfromINtheDTsameJJvirus,NNbutCCnotRBfromINdifferentJJcoronaviruses.NNSpunctamodnsubjdetdobjcasedetnmodcasedetcompoundcompoundnmodccdetamodconjcccompoundconjcasenmodcasecompoundnmodcasenmodccconjagentconjnsubjcaseamodnmodcompoundcompoundnsubjconjmarkdetnummodnsubjpassauxpassadvclcasedetamodnmodccnegcaseamodconj
displaCy
"Accumulated data suggest the importance of the compatibility of the S protein endodomain (or a cytoplasmic tailMULTI-TISSUE_STRUCTURE) and MGENE_OR_GENE_PRODUCT protein for assembly of S protein into coronavirusORGANISM or VLPsORGANISM [40, 59, 60] ; in past reports, S protein assembly occurred if these two regions are derived from the same virus, but not from different coronavirusesORGANISM.
displaCy
Accordingly,RBwePRPwereVBDsurprisedVBNtoTOfindVBtheDTefficientJJproductionNNofINchimericJJVLPsNNScarryingVBGSCoVNNSNNproteinNNandCCMHV-derivedJJN,NNMNNandCCENNproteins.NNSadvmodnsubjpassauxpassmarkxcompdetamoddobjcaseamodnmodaclamodcompounddobjccamodcompoundconjccconjconj
displaCy
Accordingly, we were surprised to find the efficient production of chimeric VLPsCELL carrying SCoV SGENE_OR_GENE_PRODUCT protein and MHV-derived N, MGENE_OR_GENE_PRODUCT and E proteins.
displaCy
TheDTpresentJJdataNNSsuggestVBPthatINtheDTendodomainNNofINSCoVNNSNNproteinNNinteractedVBDefficientlyRBwithINMHVNNMNNproteinNNandCCassembledVBNintoINVLPs.NNSdetamodnsubjmarkdetnsubjcaseamodcompoundnmodccompadvmodcasecompoundcompoundnmodccconjcasenmod
displaCy
The present data suggest that the endodomain of SCoV SGENE_OR_GENE_PRODUCT protein interacted efficiently with MHV MGENE_OR_GENE_PRODUCT protein and assembled into VLPsCELL.
displaCy
TheDTendodomainNNofINcoronavirusNNSNNproteinsNNShasVBZtwoCDpartiallyRBoverlappingVBGsub-regions,NNSaDTN-terminal,JJ\u223c18-residue-long,NNcysteine-richJJregionNNandCCaDTC-terminal,JJ\u223c27-residue-long,NNcharge-richJJregion (NNFig.NN5) .CDdetnsubjcasecompoundcompoundnmodnummodadvmodamoddobjdetamodconjamodapposccdetamodconjamodconjapposnsubj
displaCy
The endodomain of coronavirus S proteinsORGANISM has two partially overlapping sub-regions, a N-terminal, \u223c18-residue-long, cysteine-rich region and a C-terminal, \u223c27-residue-longGENE_OR_GENE_PRODUCT, charge-richCELL region (Fig. 5) .
displaCy
MHVNNlackingVBGtheDTveryRBC-terminalJJ12-amino-acidJJsegmentNNofINtheDTSNNproteinNNendodomainNNisVBZviableJJandCCreplicatesNNSwellRBinINcellNNculture,NNsuggestingVBGthatINtheDTremovalNNofIN12CDcarboxy-terminalJJresiduesNNSdoesVBZnotRBinhibitVBinteractionsNNSbetweenINtheDTmutantJJSNNproteinNNandCCtheDTMNNprotein [NN25,CD26] .CDnsubjacldetadvmodamodamoddobjcasedetcompoundcompoundnmodpredetccconjadvmodcasecompoundnmodxcompmarkdetnsubjcasenummodamodnmodauxnegccompdobjcasedetamodcompoundnmodccdetcompoundconjagentnsubj
displaCy
MHVORGANISM lacking the very C-terminal 12-amino-acid segment of the S protein endodomain is viable and replicates well in cellCELL culture, suggesting that the removal of 12 carboxy-terminal residues does not inhibit interactions between the mutant S protein and the M protein [25, 26] .
displaCy
InINcontrast,NNreplicationNNisVBZseverelyRBimpairedJJinINMHVNNmutantsNNSlackingVBGtheDTveryRBC-terminalJJ22CDaminoNNacidsNNSorCC25CDaminoNNacidsNNSofINtheDTSNNproteinNNendodomain,NNi.e.,FWconstitutingVBGtheDTmajorityNNofINtheDTcharge-richJJregion .NNcasenmodnsubjauxadvmodcasecompoundnmodacldetadvmodamodnummodcompounddobjccnummodcompoundconjcasedetcompoundcompoundnmodadvmodadvcldetdobjcasedetamodnmod
displaCy
In contrast, replication is severely impaired in MHVORGANISM mutants lacking the very C-terminal 22 amino acids or 25 amino acids of the S protein endodomain, i.e., constituting the majority of the charge-richCELL region .
displaCy
Furthermore,RBaDTsingleJJpointNNmutationNNofINaDTchargedVBNaminoNNacidNNtoTOalanineNNinINtheDTcharge-richJJregionNNofINtheDTendodomainNNhasVBZnoDTdetectableJJeffectNNonINtheDTincorporationNNofINaDTheterologousJJproteinNNcontainingVBGMHVNNSNNendodomainNNintoINvirions,NNSwhereasINmutantsNNScontainingVBGmultipleJJchargedJJresidues-toalanineNNsignificantlyRBreduceVBPtheDTincorporationNNofINtheDTproteinNNintoINvirions .NNSadvmoddetamodcompoundnsubjcasedetamodcompoundnmodcasenmodcasedetamodnmodcasedetnmodnegamoddobjcasedetnmodcasedetamodnmodaclcompoundcompounddobjcasenmodagentnsubjaclamodamoddobjadvmodmwedetdobjcasedetnmodcasenmod
displaCy
Furthermore, a single point mutation of a charged amino acidAMINO_ACID to alanineAMINO_ACID in the charge-rich region of the endodomain has no detectable effect on the incorporation of a heterologous protein containing MHV S endodomain into virions, whereas mutants containing multiple charged residues-toalanineSIMPLE_CHEMICAL significantly reduce the incorporation of the protein into virions .
displaCy
TheseDTdataNNSsuggestVBPthatINtheDTcharge-richJJregionNNofINtheDTendodomainNNplaysVBZaDTkeyJJroleNNinINtheDTselectiveJJinclusionNNofINtheDTSNNproteinNNintoINtheDTvirion.NNdetnsubjmarkdetamodnsubjcasedetnmodccompdetamoddobjcasedetamodnmodcasedetcompoundnmodcasedetnmod
displaCy
These data suggest that the charge-richCELLULAR_COMPONENT region of the endodomain plays a key role in the selective inclusion of the S protein into the virion.
displaCy
WePRPnotedVBDthatINthereEXwereVBD6CDchargedVBNresiduesNNSbetweenINtheDTtransmembraneNNdomainNNandCCtheDTC-terminalJJ12CDaminoNNacidsNNSinINbothCCSCoVNNandCCMHV,NNwhileINonlyRB3CDchargedJJresiduesNNSareVBPfoundVBNinINfelineNNinfectiousJJperitonitisNNvirus (NNFIPV) (NNFig.NN5) .CDnsubjmarkdobjccompnummodamodnegcasedetcompoundnmodccdetamodnummodcompoundconjcasenmod:npmodnmodccconjmarkadvmodnummodamodnsubjpassauxpassadvclcaseamodamodcompoundnmodapposmwensubj
displaCy
We noted that there were 6 charged residues between the transmembraneCELLULAR_COMPONENT domain and the C-terminal 12 amino acids in both SCoVSIMPLE_CHEMICAL and MHVORGANISM, while only 3 charged residues are found in feline infectious peritonitis virusORGANISM (FIPVORGANISM) (Fig. 5) .
displaCy
FIPVNNSNNproteinNNisVBZnotRBassembledVBNintoINVLPsNNSorCCvirusesNNSthatWDTcarryVBPMHVNNPM,NNNNNandCCENNproteins .NNScompoundcompoundnsubjpassauxpassnegcasenmodccconjnsubjparataxiscompounddobjapposccconjagent
displaCy
FIPV SORGANISM protein is not assembled into VLPs or viruses that carry MHV MORGANISM, N and E proteins .
displaCy
Accordingly,RBtheDTpresenceNNofINtheDTsameJJnumberNNofINchargedVBNresiduesNNSatINtheDTregionNNofINtheDTendodomain,NNwhichWDTisVBZconsideredVBNtoTObeVBimportantJJforINSNNproteinNNassemblyNNintoINvirusNNparticles,NNSinINbothCCSCoVNNandCCMHVNNimpliesVBZthatINbothDTvirusesNNShadVBDaDTsimilarJJstructureNNinINthisDTportionNNofINtheDTendodomain,NNallowingVBGthemPRPtoTOinteractVBwithINMHVNNMNNprotein.NNadvmoddetnsubjcasedetamodnmodcaseamodnmodcasedetnmodcasedetnmodnsubjpassauxpassparataxismarkpredetxcompcasecompoundcompoundnmodcasecompoundnmodcasenmod:npmodnsubjccconjmarkdetnsubjccompdetamoddobjcasedetnmodcasedetnmodadvclnsubjmarkccompcasecompoundcompoundnmod
displaCy
Accordingly, the presence of the same number of charged residues at the region of the endodomain, which is considered to be important for S protein assembly into virus particles, in both SCoVSIMPLE_CHEMICAL and MHVORGANISM implies that both viruses had a similar structure in this portion of the endodomain, allowing them to interact with MHV MORGANISM protein.
displaCy
"``InoculationNNofINmiceNNSwithINaDTmixtureNNofINchimericJJVLPsNNSandCCalum,NNbutCCnotRBthatDTofINMHVNNVLPsNNSandCCalum,NNinducedVBDantibodiesNNSthatWDTneutralizedVBDSCoV (NNFig.NN3) .CDpunctcasenmodcasedetnmodcaseamodnmodccconjccnegconjcasecompoundnmodccconjamodapposnsubjparataxisdobjapposnsubj
displaCy
"Inoculation of miceORGANISM with a mixture of chimeric VLPsCELL and alumSIMPLE_CHEMICAL, but not that of MHV VLPsORGANISM and alumSIMPLE_CHEMICAL, induced antibodies that neutralized SCoVSIMPLE_CHEMICAL (Fig. 3) .
displaCy
TheDTlackNNofINheterologousJJprotectionNNconfirmedVBDthatINtheDTSCoVNNSNNproteinNNisVBZtheDTrelevantJJconstituentNNofINtheDTchimericJJVLPs.NNSdetnsubjcaseamodnmodmarkdetamodcompoundnsubjpredetdetamodccompcasedetamodnmod
displaCy
The lack of heterologous protection confirmed that the SCoV SGENE_OR_GENE_PRODUCT protein is the relevant constituent of the chimeric VLPsCELL.
displaCy
AsINhasVBZbeenVBNobservedVBNinINotherJJVLPNNsystems [NNS61,CD62] ,CDtheDTparticlesNNSthemselvesPRPareVBPimmunogenicJJafterIN1CDorCC2CDinjectionsNNSandCCprovidedVBNprotectionNNwhenWRBchallengeNNwasVBDcarriedVBNoutRPafterINtheDTsecondJJi.m.NNmarkauxauxpasscaseamodcompoundnmodagentconjdetnsubjagentpredetconjcasenummodccconjnmodccconjdobjadvmodnsubjpassauxpassadvcladvmodcasedetamodnmod
displaCy
As has been observed in other VLPSIMPLE_CHEMICAL systems [61, 62] , the particles themselves are immunogenic after 1 or 2 injections and provided protection when challenge was carried out after the second i.m.
displaCy
injectionNNofINonlyRB2CDg.NNAdditionNNofINalumNNadjuvantNNenhancedVBDinductionNNofINneutralizingVBGantibodies,NNSandCCindeedRBtitersNNSreachedVBDthoseDTpost-infectionNNafterINtwoCDinjectionsNNSofIN2CDg.NNAllDTVLPNNgroupsNNShadVBDsignificantJJprotectionNNasINevidencedVBNbyINreductionNNinINlungNNviralJJtitersNNS2CDdaysNNSafterINvirulentJJvirusNNchallenge,NNtheDToptimumJJtimeNNtoTOdetectVBreplicationNNinINtheDTBalb/cNNmouse .NNnsubjcaseadvmodnummodcompoundnmodcaseamodnmoddobjcaseamodnmodccadvmodnsubjconjdetdobjcasenummodnmodcasenummodnmoddetcompoundnsubjconjamoddobjmarkadvclcasenmodcasecompoundamodnmodnummodnmodcaseamodcompoundnmoddetamoddobjmarkacldobjcasedetcompoundnmod
displaCy
injection of only 2 g. Addition of alumSIMPLE_CHEMICAL adjuvant enhanced induction of neutralizing antibodies, and indeed titers reached those post-infection after two injections of 2 g. All VLPCELL groups had significant protection as evidenced by reduction in lungORGAN viral titers 2 days after virulent virus challenge, the optimum time to detect replication in the Balb/c mouseORGANISM .
displaCy
HigherJJRneutralizingVBGantibodyNNtitersNNScorrelatedVBDwithINlowerJJRvirusNNcontent,NNandCCtheDT2CDgNNplusCCalumJJgroupsNNShadVBDnoDTdetectableJJvirus.NNamodamodcompoundnsubjcaseamodcompoundnmodccdetnummodnsubjcccompoundconjconjnegamoddobj
displaCy
Higher neutralizing antibody titers correlated with lower virus content, and the 2 g plus alumSIMPLE_CHEMICAL groups had no detectable virus.
displaCy
Overall,RBourPRP$dataNNSwereVBDinINagreementNNwithINpastJJreportsNNSrevealingVBGtheDTimportanceNNofINSCoV-specificJJneutralizingVBGantibodies,NNSmostJJSofINwhichWDTrecognizeVBPSNNprotein,NNforINcurbingVBGSCoVNNreplicationNNinINtheDTlung [NN21, .CDadvmodcc:preconjnsubjpredetcasecaseamodnmodacldetdobjcaseamodamodnmodnsubjcasenmodparataxiscompounddobjmarkadvclamoddobjcasedetnmodagent
displaCy
Overall, our data were in agreement with past reports revealing the importance of SCoV-specific neutralizing antibodies, most of which recognize S protein, for curbing SCoVSIMPLE_CHEMICAL replication in the lungTISSUE [21, .
displaCy
"``ImmunizationNNofINmiceNNSwithINVenezuelanJJequineJJencephalitisNNvirusNNrepliconJJparticlesNNSencodingVBGSCoVNNNNNproteinNNinducedVBDanDTenhancedVBNimmunopathology,NNthatWDTincludedVBDinfiltratingVBGeosinophilsNNSinINtheDTlungs,NNSfollowingVBGSCoVNNchallenge ,NNimplyingVBGtheDTpossibilityNNthatINaDTSCoVNNvaccineNNlackingVBGNNNproteinNNexpressionNNmayMDbeVBdesirable,JJasINitPRPwouldMDreduceVBtheDTpotentialJJriskNNofINimmunopathologicalJJchangesNNSprimedVBNbyINimmunization.NNpunctnsubjcasenmodcasecompoundcompoundcompoundcompoundcompoundnmodaclamodcompounddobjdetamoddobjnsubjagentamoddobjcasedetnmodcaseamodnmodadvcldetdobjmarkdetamodnsubjaclcompoundcompounddobjauxpredetccompmarknsubjauxadvcldetamoddobjcaseamodnmodaclcasenmod
displaCy
"Immunization of miceORGANISM with Venezuelan equine encephalitis virus replicon particles encodingORGANISM SCoV NGENE_OR_GENE_PRODUCT protein induced an enhanced immunopathology, that included infiltrating eosinophilsCELL in the lungsORGAN, following SCoVSIMPLE_CHEMICAL challenge , implying the possibility that a SCoV vaccineSIMPLE_CHEMICAL lacking N protein expression may be desirable, as it would reduce the potential risk of immunopathological changes primed by immunization.
displaCy
InINthisDTregard,NNchimericJJVLPsNNScarryingVBGMHVNNNNNproteinNNorCCthoseDTlackingVBGNNNprotein (NNunpublishedJJdata)NNScouldMDeliminateVBtheDTpossibleJJinductionNNofINSCoVNNNNNprotein-inducedJJimmunopathology.NNcasedetnmodamodnsubjaclcompoundcompounddobjccconjaclcompounddobjamodapposauxdetamoddobjcaseamodcompoundamodnmod
displaCy
In this regard, chimeric VLPsCELL carrying MHV N proteinORGANISM or those lacking N protein (unpublished data) could eliminate the possible induction of SCoV N protein-induced immunopathologySIMPLE_CHEMICAL.
displaCy
TheDTefficientJJproductionNNofINchimericJJVLPNNfromINCHONNcellsNNSasINwePRPreportVBPinINthisDTstudyNNisVBZpromisingJJasINaDTnewJJstrategyNNforINpreparingVBGVLP-basedJJSCoVNNvaccineNNfromINmammalianJJcells;NNSitsPRP$suitabilityNNforINpreparingVBGlargeJJquantitiesNNSofINVLP-basedJJvaccineNNagainstINSCoVNNremainsVBZtoTObeVBdetermined.VBNdetamodnsubjcaseamodnmodcasecompoundnmodmarknsubjnmodcasedetnmodauxcasedetamodnmodmarkaclamodamoddobjcaseamodnmodcc:preconjnsubjpreppcompamoddobjcaseamodnmodcasenmodmwemarkauxpassxcomp
displaCy
The efficient production of chimeric VLPCELL from CHO cellsCELL as we report in this study is promising as a new strategy for preparing VLP-based SCoV vaccine from mammalian cellsCELL; its suitability for preparing large quantities of VLP-based vaccine against SCoVSIMPLE_CHEMICAL remains to be determined.
displaCy
FurtherJJimprovementsNNSofINchimericJJVLPNNproduction,NNe.g.,FWcodon-optimizationNNofINtheDTSCoVNNSNNgeneNNandCCMHVNNN,NNMNNandCCENNgenesNNSinINtheDTexpressionNNplasmidsNNSandCCdevelopmentNNofINanDTinexpensiveJJtransfectionNNreagentNNand/orCCprocedure,NNwouldMDbeVBrequiredVBNforINtheDTsubstantialJJenhancementNNofINchimericJJVLPNNproduction.NNamodnsubjpasscaseamodcompoundnmodadvmodnmodcasedetamodcompoundnmodcccompoundconjagentcccompoundconjcasedetcompoundnmodccconjcasedetamodcompoundnmodccconjauxauxpasscasedetamodnmodcaseamodcompoundnmod
displaCy
Further improvements of chimeric VLPCELL production, e.g., codon-optimization of the SCoV SGENE_OR_GENE_PRODUCT gene and MHV N, M andORGANISM EGENE_OR_GENE_PRODUCT genes in the expression plasmids and development of an inexpensive transfection reagent and/or procedure, would be required for the substantial enhancement of chimeric VLPCELL production.
displaCy
"``AccumulatedJJdataNNSsuggestVBPthatINantibodiesNNSagainstINtheDTFIPVNNSNNproteinNNfailVBPtoTOprotectVBcatsNNSfromINFIPVNNchallengeNNandCCenhanceVBPvirusNNreplicationNNinINtheDThostNNthroughINantibody-dependentJJenhancement (NNADE) .NNpunctamodnsubjmarknsubjcasedetcompoundcompoundnmodccompmarkxcompdobjcasecompoundnmodccconjcompounddobjcasedetnmodcaseamodnmodappos
displaCy
"Accumulated data suggest that antibodies against the FIPV SORGANISM protein fail to protect catsORGANISM from FIPVORGANISM challenge and enhance virus replication in the host through antibody-dependent enhancement (ADE) .
displaCy
ItPRPhasVBZbeenVBNreportedVBNthatINantibodiesNNSthatWDTneutralizeVBPmostRBShumanJJSCoVNNisolatesNNSenhanceVBPentryNNofINaDTSCoVNNisolateNNfromINtheDTcivetNNinINtheDTcellNNcultureNNlevel ,NNbutCCitPRPisVBZunclearJJwhetherINADENNoccursVBZinINanimalsNNSthatWDTareVBPimmunizedVBNwithINSCoVNNvaccineNNcandidates.NNSnsubjpassauxauxpassmarknsubjnsubjparataxisamodamodamoddobjccompdobjcasedetamodaclcasedetnmodcasedetcompoundcompoundnmodccnsubjpredetconjmarknsubjccompcasenmodnsubjpassauxpassparataxiscaseamodcompoundnmod
displaCy
It has been reported that antibodies that neutralize most human SCoV isolatesORGANISM enhance entry of a SCoVSIMPLE_CHEMICAL isolate from the civet in the cellCELL culture level , but it is unclear whether ADE occurs in animals that are immunized with SCoV vaccineSIMPLE_CHEMICAL candidates.
displaCy
WePRPdidVBDnotRBobserveVBsignsNNSofINADENNinINourPRP$study;NNtheDTmiceNNSthatWDTwereVBDimmunizedVBNwithINchimericJJVLPsNNSorCCinitiallyRBinoculatedVBNwithINSCoVNNefficientlyRBsuppressedVBDchallengedVBNSCoVNNreplication.NNnsubjauxnegdobjcasenmodcasecc:preconjnmoddetnsubjnsubjpassauxpassparataxiscaseamodnmodccadvmodconjcasenmodadvmodmwexcompamoddobj
displaCy
We did not observe signs of ADE in our study; the miceORGANISM that were immunized with chimeric VLPs or initially inoculated with SCoVSIMPLE_CHEMICAL efficiently suppressed challenged SCoVSIMPLE_CHEMICAL replication.
displaCy
SNNproteinNNinINchimericJJVLPsNNSwasVBDderivedVBNfromINaDTSCoVNNUrbaniNNstrain,NNwhichWDTwasVBDalsoRBusedVBNasINtheDTchallengeNNvirus,NNandCCVLPsNNSinducedVBDantibodiesNNSthatWDTreadilyRBneutralizedVBDSCoVNNinFWvitro.FWcompoundnsubjpasscaseamodnmodauxpasscasedetamodcompoundnmodnsubjpassauxpassadvmodparataxiscasedetcompoundnmodccnsubjconjdobjnsubjadvmodparataxisdobjcompoundadvmod
displaCy
S protein in chimeric VLPsCELL was derived from a SCoV UrbaniORGANISM strain, which was also used as the challenge virus, and VLPsCELL induced antibodies that readily neutralized SCoVSIMPLE_CHEMICAL in vitro.
displaCy
InINthisDTregard,NNitPRPisVBZworthJJnotingVBGthatINimmunizationNNofINmiceNNSwithINaDTVenezuelanJJequineJJencephalitisNNvirusNNrepliconNNcarryingVBGSNNproteinNNofINtheDTUrbaniNNstrainNNshowedVBDonlyRBlimitedJJprotectionNNagainstINheterologousJJSCoVNNthatWDTcontainedVBDSNNgeneNNfromINtheDThumanJJGD03NNisolate .NNcasedetnmodnsubjpredetxcompmarknsubjcasenmodcasedetcompoundcompoundcompoundcompoundnmodaclcompounddobjcasedetcompoundnmodccompadvmodamoddobjcaseamodnmodnsubjparataxiscompounddobjcasedetamodcompoundnmod
displaCy
In this regard, it is worth noting that immunization of miceORGANISM with a Venezuelan equine encephalitis virus replicon carrying S protein of the Urbani strainORGANISM showed only limited protection against heterologous SCoVSIMPLE_CHEMICAL that contained S gene from the human GD03ORGANISM isolate .
displaCy
"``SubbaraoNNPetFWal.FWreportedVBDmildJJandCCfocalJJperibronchiolarJJmononuclearJJinflammatoryJJinfiltratesNNSandCCSCoVNNantigensNNSinINbronchiolarJJepithelialJJcellsNNSinINtheDTlungsNNSofINBALB/cNNmiceNNSatIN2CDdaysNNSpostVBPSCoVNNinoculation .NNpunctnsubjagentagentamodccconjcompoundamodamoddobjccamodconjcaseamodamodnmodcasedetnmodcasecompoundnmodcasenummodcompoundadvclamoddobj
displaCy
"Subbarao et al. reported mild and focal peribronchiolar mononuclearCELL inflammatory infiltratesTISSUE and SCoV antigensGENE_OR_GENE_PRODUCT in bronchiolar epithelial cellsCELL in the lungsORGAN of BALB/c miceORGANISM at 2 days post SCoVSIMPLE_CHEMICAL inoculation .
displaCy
StainingVBGwithINSCoVJJmonoclonalJJantibodyNNtoTONNNprotein,NNwePRPalsoRBfoundVBDSCoVNNantigenNNwithinINbronchiolarJJepithelialJJcellsNNSafterINchallengeNNofINunimmunizedJJmice.NNSadvclcaseamodamodnmodcasecompoundnmodnsubjadvmodamoddobjcaseamodamodnmodcasenmodcaseamodnmod
displaCy
Staining with SCoV monoclonal antibodyGENE_OR_GENE_PRODUCT to N protein, we also found SCoV antigenGENE_OR_GENE_PRODUCT within bronchiolar epithelial cellsCELL after challenge of unimmunized miceORGANISM.
displaCy
ConsistentJJwithINtheDTSCoVNNtitersNNSinINtheDTlungs,NNSnoDTviralJJantigenNNwasVBDdetectedVBNinINtheDTlungsNNSofINtheDTmiceNNSinoculatedVBNwithINchimericJJVLPs.NNSadvclcasedetamodnmodcasedetnmodnegamodnsubjpassauxpasscasedetnmodcasedetnmodaclcaseamodnmod
displaCy
Consistent with the SCoVSIMPLE_CHEMICAL titers in the lungsORGAN, no viral antigen was detected in the lungsORGAN of the miceORGANISM inoculated with chimeric VLPsCELL.
displaCy
TheDTabsenceNNofINSCoVNNantigenNNinINtheDTlungsNNSofINVLP-immunizedJJmiceNNSorCCmiceNNSpre-exposedVBDtoTOliveVBvirus,NNasINindicatedVBNbyINtheDTresultsNNSofINIHCNNstaining (NNFig.NN4B) ,NNstronglyRBargueVBPforINtheDTefficacyNNofINchimericJJVLPsNNSinINprotectingVBGagainstINSARS-CoVJJinfection.NNdetnsubjcaseamodnmodcasedetnmodcaseamodnmodccconjcaseamodnmodmarkadvclcasedetnmodcasecompoundnmodcompoundapposadvmodmwecasedetnmodcaseamodnmodmarkadvclcaseamodnmod
displaCy
The absence of SCoV antigenGENE_OR_GENE_PRODUCT in the lungsORGAN of VLP-immunized miceORGANISM or miceORGANISM pre-exposed to live virus, as indicated by the results of IHCGENE_OR_GENE_PRODUCT staining (Fig. 4B) , strongly argue for the efficacy of chimeric VLPsCELL in protecting against SARS-CoVORGANISM infection.
displaCy
SCoV-inducedJJhistopathologicalJJchangesNNSthatWDTwereVBDdetectedVBNinINunimmunizedJJmiceNNSinINourPRP$studyNNappearedVBDtoTObeVBsimilarJJorCCslightlyRBmoreRBRsevere,JJespeciallyRBwithINrespectNNtoTOtheDTchangesNNSinINtheDTbronchioles,NNSthanINthoseDTdescribedVBNinINtheDTreportNNofINSubbaraoNNPetFWal. .FWamodamodnsubjnsubjpassauxpassparataxiscaseamodnmodcasecc:preconjnmodmarkpredetxcompccadvmodadvmodconjadvmodcasenmodcasedetnmodcasedetnmodcasenmodaclcasedetnmodcasenmodagentagent
displaCy
SCoV-induced histopathological changes that were detected in unimmunized miceORGANISM in our study appeared to be similar or slightly more severe, especially with respect to the changes in the bronchiolesTISSUE, than those described in the report of Subbarao et al. .
displaCy
AfterINSCoVNNchallenge,NNtheDTmiceNNSimmunizedVBNwithINchimericJJVLPsNNSshowedVBDmixedJJperibronchiolarJJinflammatoryJJinfiltratesNNSandCCaDTslightJJthickeningNNofINtheDTperibronchiolarJJinterstitiumNNandCCalveolarJJwalls.NNScaseamodnmoddetnsubjaclcaseamodnmodamodcompoundamoddobjccdetamodconjcasedetcompoundnmodccamodconj
displaCy
After SCoVSIMPLE_CHEMICAL challenge, the miceORGANISM immunized with chimeric VLPsCELL showed mixed peribronchiolar inflammatory infiltratesTISSUE and a slight thickening of the peribronchiolar interstitiumMULTI-TISSUE_STRUCTURE and alveolar wallsTISSUE.
displaCy
TheseDTlesionsNNSinINimmunizedJJmiceNNSwereVBDmilderJJthanINseenVBNinINunimmunizedJJmice,NNSdemonstratingVBGtheDTeffectsNNSofINchimericJJVLPNNimmunizationNNinINsuppressingVBGSCoVinducedJJcytopathologicalJJchangesNNSinINtheDTlungs.NNSdetnsubjcaseamodnmodpredetmarkadvclcaseamodnmodxcompdetdobjcaseamodcompoundnmodmarkaclamodamoddobjcasedetnmod
displaCy
These lesionsPATHOLOGICAL_FORMATION in immunized miceORGANISM were milder than seen in unimmunized miceORGANISM, demonstrating the effects of chimeric VLPCELL immunization in suppressing SCoVinduced cytopathological changes in the lungsORGAN.
displaCy
TheDTmiceNNSimmunizedVBNwithINaDTmixtureNNofINchimericJJVLPNNandCCalumNNhadVBDhigherJJRneutralizingVBGantibodyNNtitersNNSthanINdidVBDthoseDTimmunizedVBNwithINaDTmixtureNNofINtheDTchimericJJVLPNNandCCPBS,NNwhileINbothDTgroupsNNSofINtheDTmiceNNSshowedVBDsimilarJJlevelsNNSofINSCoV-inducedJJlungNNcytopathology,NNsuggestingVBGneitherCCalumNNnorCCneutralizingVBGantibodyNNtitersNNSwereVBDsoleJJdeterminantNNofINtheDTseverityNNofINinflammatoryJJresponses.NNSdetnsubjaclcasedetnmodcaseamodnmodccconjamodamodcompounddobjmarkadvclnegaclcasedetnmodcasedetamodnmodccconjmarkdetnsubjcasedetnmodadvclamoddobjcaseamodcompoundnmodadvclnmod:npmodnsubjccconjcompoundconjpredetamoddobjcasedetnmodcaseamodnmod
displaCy
The miceORGANISM immunized with a mixture of chimeric VLPCELL and alumSIMPLE_CHEMICAL had higher neutralizing antibody titers than did those immunized with a mixture of the chimeric VLPCELL and PBS, while both groups of the miceORGANISM showed similar levels of SCoV-induced lung cytopathologyCANCER, suggesting neither alumSIMPLE_CHEMICAL nor neutralizing antibody titers were sole determinant of the severity of inflammatory responses.
displaCy
PossibleJJcellularJJimmuneJJresponsesNNSagainstINchimericJJVLPNNproteinsNNSwereVBDsuggestedVBNbyINtheDTobservationNNthatINtheDTmiceNNSinoculatedVBNwithINaDTmixtureNNofINchimericJJVLPsNNSandCCPBSNNandCCthatDTofINchimericJJVLPsNNSandCCalum,NNbutCCnotRBotherJJgroupsNNSofINmice,NNSshowedVBDinfiltrationNNofINneutrophilsNNSandCCeosinophilsNNSaroundINtheDTbronchiolesNNSandCCbloodNNvessels.NNSamodamodamodnsubjpasscaseamodcompoundnmodauxpasscasedetnmodmarkdetnsubjaclcasedetnmodcaseamodnmodccconjccconjcaseamodnmodccconjccnegamodconjcasenmodccompdobjcasenmodccconjcasedetnmodcccompoundconj
displaCy
Possible cellularCELL immune responses against chimeric VLPCELL proteins were suggested by the observation that the miceORGANISM inoculated with a mixture of chimeric VLPsCELL and PBS and that of chimeric VLPsCELL and alumSIMPLE_CHEMICAL, but not other groups of miceORGANISM, showed infiltration of neutrophilsCELL and eosinophilsCELL around the bronchiolesTISSUE and blood vesselsMULTI-TISSUE_STRUCTURE.
displaCy
InINcontrast,NNitPRPhasVBZbeenVBNreportedVBNthatINimmunizationNNofINmiceNNSwithINaDTmixtureNNofINinactivatedVBNSCoVNNwithINadjuvantJJMF59NNprimaryJJelicitedVBDhumoralJJimmuneJJresponse .NNcasenmodnsubjpassauxauxpassmarkccompcasenmodcasedetnmodcaseamodnmodcaseamodpreconjamodamodamodamodnmod
displaCy
In contrast, it has been reported that immunization of miceORGANISM with a mixture of inactivated SCoVSIMPLE_CHEMICAL with adjuvant MF59SIMPLE_CHEMICAL primary elicited humoral immune response .
displaCy
FurtherJJinvestigation,NNusingVBGnotRBonlyRBBalb/cNNmiceNNSbutCCalsoRBotherJJSCoVNNsusceptibleJJmice [NNS41,CD76,CD77]CDandCCotherJJanimalNNmodels [NNS78,CD79] ,CDwillMDbeVBnecessaryJJtoTOevaluateVBtheDTimplicationsNNSandCCimpactNNofINcellularJJimmuneJJactivities,NNSincludingVBGpulmonaryJJinfiltrationNNofINneutrophilsNNSandCCeosinophils,NNSasRBwellRBasINtheDTutilityNNofINchimericJJVLPsNNSasINaDTSCoVNNvaccine.NNamodnsubjaclnegnmod:npmodcompounddobjccadvmodamodamodamodconjagentconjconjccamodcompoundconjagentconjauxpredetmarkxcompdetdobjccconjcaseamodamodnmodcaseamodnmodcasenmodccconjccquantmodquantmoddetconjcaseamodnmodcasedetamodnmod
displaCy
Further investigation, using not only Balb/c miceORGANISM but also other SCoVSIMPLE_CHEMICAL susceptible miceORGANISM [41, 76, 77] and other animal models [78, 79] , will be necessary to evaluate the implications and impact of cellularCELL immune activities, including pulmonaryORGAN infiltration of neutrophilsCELL and eosinophilsCELL, as well as the utility of chimeric VLPsCELL as a SCoV vaccineSIMPLE_CHEMICAL.
displaCy
"``EvolutionaryJJinsightsNNSintoINtheDTecologyNNofINcoronaviruses",NNS_SPpunctamodcasedetnmodpreppunct
displaCy
"Evolutionary insights into the ecology of coronavirusesORGANISM",
displaCy
"``Cross-hostJJevolutionNNofINsevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNNinINpalmNNcivetNNandCChuman",JJ_SPpunctamodcaseamodamodamodcompoundnmodprepcompoundpobjccpunct
displaCy
"Cross-hostCELL evolution of severe acute respiratory syndrome coronavirusORGANISM in palm civetORGANISM_SUBDIVISION and humanORGANISM",
displaCy
"``TheDTmolecularJJbiologyNNofINcoronaviruses",NNS_SPpunctdetamodcasenmod
displaCy
"The molecular biology of coronavirusesORGANISM",
displaCy
"``ReceptorNNandCCviralJJdeterminantsNNSofINSARS-coronavirusJJadaptationNNtoTOhumanJJACE2",NN_SPpunctcompoundccconjcaseamodnmodcaseamodnmod
displaCy
"Receptor and viral determinants of SARS-coronavirusSIMPLE_CHEMICAL adaptation to human ACE2ORGANISM",
displaCy
"``AminoNNacidNNsubstitutionsNNSandCCanDTinsertionNNinINtheDTspikeNNglycoproteinNNextendVBPtheDThostNNrangeNNofINtheDTmurineJJcoronavirusNNMHV-A59",NN_SPpunctcompoundcompoundnsubjccdetconjcasedetamodnmoddetcompounddobjcasedetamodcompoundnmod
displaCy
"Amino acidAMINO_ACID substitutions and an insertion in the spike glycoproteinGENE_OR_GENE_PRODUCT extend the host range of the murine coronavirus MHV-A59ORGANISM",
displaCy
"``MurineJJcoronavirus-inducedJJhepatitis:NNJHMNNgeneticJJbackgroundNNeliminatesVBZA59NNPspike-determinedJJhepatotropism",NN_SPpunctamodamodnsubjcompoundamodnsubjcompoundamoddobj
displaCy
"Murine coronavirus-induced hepatitisORGANISM: JHMORGANISM genetic background eliminates A59GENE_OR_GENE_PRODUCT spike-determined hepatotropism",
displaCy
"``RecombinantJJavianJJinfectiousJJbronchitisNNvirusNNexpressingVBGaDTheterologousJJspikeNNgeneNNdemonstratesVBZthatINtheDTspikeJJproteinNNisVBZaDTdeterminantNNofINcellNNtropism",NN_SPpunctamodamodamodcompoundnsubjacldetamodamoddobjmarkdetamodnsubjpredetdetccompcasecompoundnmod
displaCy
"Recombinant avian infectious bronchitis virus expressing a heterologous spike gene demonstrates that the spike protein is a determinant of cellCELL tropism",
displaCy
"``CoronavirusNNspikeNNproteinsNNSinINviralJJentryNNandCCpathogenesis",NN_SPpunctcompoundamodcaseamodnmodccpunct
displaCy
"CoronavirusORGANISM spike proteins in viral entry and pathogenesis",
displaCy
"``TargetedJJrecombinationNNdemonstratesVBZthatINtheDTspikeNNgeneNNofINtransmissibleJJgastroenteritisNNcoronavirusNNisVBZaDTdeterminantNNofINitsPRP$entericJJtropismNNandCCvirulence",NN_SPpunctamodnsubjmarkdetamodnsubjcaseamodcompoundnmodpredetdetccompcasecc:preconjamodnmodccpunct
displaCy
"Targeted recombination demonstrates that the spike gene of transmissible gastroenteritis coronavirusORGANISM is a determinant of its enteric tropism and virulence",
displaCy
"``PathogenesisNNofINchimericJJMHV4/MHV-A59JJrecombinantJJviruses:NNStheDTmurineJJcoronavirusNNspikeNNproteinNNisVBZaDTmajorJJdeterminantNNofINneurovirulence",NN_SPpunctcaseamodcompoundamodnmoddetamodcompoundamodnsubjpredetdetamodagentcasenmod
displaCy
"Pathogenesis of chimeric MHV4/MHV-A59 recombinant virusesGENE_OR_GENE_PRODUCT: the murine coronavirusORGANISM spike protein is a major determinant of neurovirulenceORGANISM",
displaCy
"``B-cellNNresponsesNNSinINpatientsNNSwhoWPhaveVBPrecoveredVBNfromINsevereJJacuteJJrespiratoryJJsyndromeNNtargetNNaDTdominantJJsiteNNinINtheDTS2NNdomainNNofINtheDTsurfaceNNspikeNNglycoprotein",NN_SPpunctcompoundnsubjcasenmodnsubjauxparataxiscaseamodamodamodnmoddetamoddobjcasedetcompoundnmodcasedetcompoundamodpunct
displaCy
"B-cellCELL responses in patientsORGANISM who have recovered from severe acute respiratory syndrome target a dominant site in the S2 domain of the surface spike glycoprotein",
displaCy
"``IdentificationNNofINtwoCDneutralizingVBGregionsNNSonINtheDTsevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNNspikeNNglycoproteinNNproducedVBNfromINtheDTmammalianJJexpressionNNsystem",NN_SPpunctcasenummodamodnmodcasedetamodamodamodcompoundcompoundamodnmodaclcasedetamodcompoundnmod
displaCy
"Identification of two neutralizing regions on the severe acute respiratory syndrome coronavirusORGANISM spike glycoprotein produced from the mammalian expression system",
displaCy
"``AminoNNacidsNNS1055CDtoTO1192CDinINtheDTS2NNregionNNofINsevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNNSNNproteinNNinduceVBPneutralizingVBGantibodies:NNSimplicationsNNSforINtheDTdevelopmentNNofINvaccinesNNSandCCantiviralJJagents",NNS_SPpunctcompoundnsubjnsubjexplnsubjcasedetcompoundnmodcaseamodamodamodcompoundcompoundcompoundnmodamoddobjagentcasedetnmodcasenmodccamodpunct
displaCy
"Amino acids 1055AMINO_ACID to 1192 in the S2 region of severe acute respiratory syndrome coronavirus S protein induce neutralizing antibodies: implications for the development of vaccines and antiviral agents",
displaCy
ADTDNANNvaccineNNinducesVBZSARSNNPcoronavirusNNneutralizationNNandCCprotectiveJJimmunityNNinINmice",NNS_SPdetcompoundnsubjcompoundcompounddobjccamodconjcasenmod
displaCy
A DNACELLULAR_COMPONENT vaccine induces SARS coronavirusORGANISM neutralization and protective immunity in miceORGANISM",
displaCy
"``SevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNNspikeNNproteinNNexpressedVBNbyINattenuatedVBNvacciniaNNvirusNNprotectivelyNNimmunizesVBZmice",NNS_SPpunctamodamodamodcompoundcompoundamodnsubjaclcaseamodcompoundcompoundnmoddobj
displaCy
"Severe acute respiratory syndrome coronavirusORGANISM spike protein expressed by attenuated vaccinia virusORGANISM protectively immunizes miceORGANISM",
displaCy
"''ContributionsNNSofINtheDTstructuralJJproteinsNNSofINsevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNNtoTOprotectiveJJimmunity",NN_SPpunctcasedetamodnmodcaseamodamodamodcompoundnmodcaseamodnmod
displaCy
"Contributions of the structural proteins of severe acute respiratory syndrome coronavirusORGANISM to protective immunity",
displaCy
"``EvaluationNNofINhumanJJmonoclonalJJantibodyNN80RCDforINimmunoprophylaxisNNofINsevereJJacuteJJrespiratoryJJsyndromeNNbyINanDTanimalNNstudy,NNepitopeNNmapping,NNandCCanalysisNNofINspikeJJvariants",NNS_SPpunctcaseamodamodnmodagentcasenmodcaseamodamodamodnmodcasedetcompoundnmodcompoundconjccconjcaseamodnmod
displaCy
"Evaluation of human monoclonal antibodyORGANISM 80R for immunoprophylaxis of severe acute respiratory syndrome by an animal study, epitope mapping, and analysis of spike variants",
displaCy
"``DevelopmentNNandCCcharacterizationNNofINaDTsevereJJacuteJJrespiratoryJJsyndrome-associatedJJcoronavirus-neutralizingJJhumanJJmonoclonalJJantibodyNNthatWDTprovidesVBZeffectiveJJimmunoprophylaxisNNinINmice",NNS_SPpunctccconjcasedetamodamodamodamodamodamodamodnmodnsubjparataxisamoddobjcasepunct
displaCy
"Development and characterization of a severe acute respiratory syndrome-associated coronavirus-neutralizing human monoclonal antibodyORGANISM that provides effective immunoprophylaxis in miceORGANISM",
displaCy
"``PotentJJneutralizationNNofINsevereJJacuteJJrespiratoryJJsyndrome (NNSARS)NNcoronavirusNNbyINaDThumanJJmAbNNtoTOS1NNproteinNNthatWDTblocksVBZreceptorNNassociation",NN_SPpunctamodcaseamodamodamodcompoundcompoundnmodcasedetamodnmodcasecompoundnmodnsubjparataxiscompoundpunct
displaCy
"Potent neutralization of severe acute respiratory syndrome (SARS) coronavirusORGANISM by a human mAbORGANISM to S1 protein that blocks receptor association",
displaCy
"``PriorJJinfectionNNandCCpassiveJJtransferNNofINneutralizingVBGantibodyNNpreventVBPreplicationNNofINsevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNNinINtheDTrespiratoryJJtractNNofINmice",NNS_SPpunctamodnsubjccamodconjcaseamodnmoddobjcaseamodamodamodcompoundnmodcasedetamodnmodcasepunct
displaCy
"Prior infection and passive transfer of neutralizing antibody prevent replication of severe acute respiratory syndrome coronavirusORGANISM in the respiratory tract of miceORGANISM",
displaCy
"``Nucleocapsid-independentJJassemblyNNofINcoronavirus-likeJJparticlesNNSbyINco-expressionNNofINviralJJenvelopeNNproteinNNgenes",NNS_SPpunctamodcaseamodnmodcasenmodcaseamodcompoundcompoundpunct
displaCy
"Nucleocapsid-independent assembly of coronavirus-like particlesGENE_OR_GENE_PRODUCT by co-expression of viral envelope protein genes",
displaCy
"``GeneticJJanalysisNNofINdeterminantsNNSforINspikeJJglycoproteinNNassemblyNNintoINmurineJJcoronavirusNNvirions:NNSdistinctJJrolesNNSforINcharge-richJJandCCcysteine-richJJregionsNNSofINtheDTendodomain",NN_SPpunctamodcasenmodcaseamodcompoundnmodcaseamodcompoundnmodamodagentcaseamodccconjnmodcasedetpunct
displaCy
"Genetic analysis of determinants for spike glycoprotein assembly into murine coronavirus virionsORGANISM: distinct roles for charge-richCELL and cysteine-rich regions of the endodomain",
displaCy
"``MappingNNofINtheDTcoronavirusNNmembraneNNproteinNNdomainsNNSinvolvedVBNinINinteractionNNwithINtheDTspikeNNprotein",NN_SPpunctcasedetcompoundcompoundcompoundnmodaclcasenmodcasedetamodpunct
displaCy
"Mapping of the coronavirus membrane protein domainsORGANISM involved in interaction with the spike protein",
displaCy
"``ProteinNNinteractionsNNSduringINcoronavirusNNassembly",NN_SPpunctcompoundcasecompoundnmod
displaCy
"Protein interactions during coronavirusORGANISM assembly",
displaCy
"``CooperationNNofINanDTRNANNpackagingNNsignalNNandCCaDTviralJJenvelopeNNproteinNNinINcoronavirusNNRNANNpackaging",NN_SPpunctcasedetcompoundcompoundnmodccdetamodcompoundconjcasecompoundcompoundnmod
displaCy
"Cooperation of an RNA packaging signal and a viral envelope protein in coronavirusORGANISM RNA packaging",
displaCy
"``GeneticJJevidenceNNforINaDTstructuralJJinteractionNNbetweenINtheDTcarboxyNNterminiNNSofINtheDTmembraneNNandCCnucleocapsidNNproteinsNNSofINmouseNNhepatitisNNvirus",NN_SPpunctamodcasedetamodnmodcasedetcompoundnmodcaseexplnmodccconjagentcasecompoundcompoundpunct
displaCy
"Genetic evidence for a structural interaction between the carboxy termini of the membraneCELLULAR_COMPONENT and nucleocapsidGENE_OR_GENE_PRODUCT proteins of mouse hepatitis virusORGANISM",
displaCy
"``CharacterizationNNofINtheDTcoronavirusNNMNNproteinNNandCCnucleocapsidNNinteractionNNinINinfectedJJcells",NNS_SPpunctcasedetcompoundcompoundnmodcccompoundconjcaseamodnmod
displaCy
"Characterization of the coronavirus MORGANISM protein and nucleocapsidGENE_OR_GENE_PRODUCT interaction in infected cellsCELL",
displaCy
"``IsolationNNofINcoronavirusNNenvelopeNNglycoproteinsNNSandCCinteractionNNwithINtheDTviralJJnucleocapsid",NN_SPpunctcasecompoundcompoundnmodccconjcasedetamodnmod
displaCy
"Isolation of coronavirus envelope glycoproteinsORGANISM and interaction with the viral nucleocapsidORGANISM",
displaCy
"``NucleocapsidindependentJJspecificJJviralJJRNANNpackagingNNviaINviralJJenvelopeNNproteinNNandCCviralJJRNANNsignal",NN_SPpunctcompoundamodamodcompoundcaseamodcompoundnmodccamodcompoundpunct
displaCy
"NucleocapsidindependentSIMPLE_CHEMICAL specific viral RNA packaging via viral envelope protein and viral RNA signal",
displaCy
"``ReleaseNNofINcoronavirusNNENNproteinNNinINmembraneNNvesiclesNNSfromINvirus-infectedJJcellsNNSandCCENNproteinexpressingJJcells",NNS_SPpunctcasecompoundcompoundnmodcasecompoundnmodcaseamodnmodcccompoundcompoundpunct
displaCy
"Release of coronavirus EORGANISM protein in membrane vesiclesCELLULAR_COMPONENT from virus-infected cellsCELL and E proteinexpressing cellsCELL",
displaCy
"``CoronavirusNNpseudoparticlesNNSformedVBNwithINrecombinantJJMNNandCCENNproteinsNNSinduceVBPalphaNNinterferonNNsynthesisNNbyINleukocytes",NNS_SPpunctcompoundnsubjaclcaseamodnmodcccompoundconjcompoundcompounddobjcasenmod
displaCy
"Coronavirus pseudoparticlesORGANISM formed with recombinant M and E proteinsGENE_OR_GENE_PRODUCT induce alpha interferonGENE_OR_GENE_PRODUCT synthesis by leukocytesCELL",
displaCy
"``SevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNN7aNNaccessoryNNproteinNNisVBZaDTviralJJstructuralJJprotein",NN_SPpunctamodamodamodcompoundcompoundcompoundcompoundnsubjpredetdetamodamod
displaCy
"Severe acute respiratory syndrome coronavirus 7a accessoryORGANISM protein is a viral structural protein",
displaCy
"``EfficientJJassemblyNNandCCreleaseNNofINSARSNNPcoronavirus-likeJJparticlesNNSbyINaDTheterologousJJexpressionNNsystem",NN_SPpunctcompoundccconjcasecompoundamodnmodcasedetamodcompoundnmod
displaCy
"Efficient assembly and release of SARS coronavirus-like particlesORGANISM by a heterologous expression system",
displaCy
"``ImmuneJJresponsesNNSagainstINsevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNNinducedVBNbyINvirus-likeJJparticlesNNSinINmice",NNS_SPpunctamodcaseamodamodamodcompoundnmodaclcaseamodnmodcasepunct
displaCy
"ImmuneCELL responses against severe acute respiratory syndrome coronavirusORGANISM induced by virus-like particlesORGANISM in miceORGANISM",
displaCy
"``GenerationNNofINsyntheticJJsevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNNpseudoparticles:NNSimplicationsNNSforINassemblyNNandCCvaccineNNproduction",NN_SPpunctcaseamodamodamodamodcompoundcompoundnmodagentcasenmodcccompoundconj
displaCy
"Generation of synthetic severe acute respiratory syndrome coronavirus pseudoparticlesORGANISM: implications for assembly and vaccine production",
displaCy
"``RetargetingNNofINcoronavirusNNbyINsubstitutionNNofINtheDTspikeNNglycoproteinNNectodomain:NNcrossingVBGtheDThostNNcellNNspeciesNNSbarrier",NN_SPpunctcasenmodcasenmodcasedetamodcompoundnmodacldetcompoundcompoundcompounddobj
displaCy
"Retargeting of coronavirusORGANISM by substitution of the spike glycoprotein ectodomain: crossing the host cellCELL species barrier",
displaCy
"``SevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNNinfectionNNofINmiceNNStransgenicJJforINtheDThumanJJAngiotensin-convertingJJenzymeNN2CDvirusNNreceptor",NN_SPpunctamodamodamodcompoundcompoundcasenmodnummodcasedetamodamodcompoundnummodcompoundpunct
displaCy
"Severe acute respiratory syndrome coronavirusORGANISM infection of miceORGANISM transgenic for the human Angiotensin-converting enzyme 2 virus receptorORGANISM",
displaCy
"``EnvelopeNNglycoproteinNNinteractionsNNSinINcoronavirusNNassembly",NN_SPpunctcompoundcompoundcasecompoundnmod
displaCy
"Envelope glycoprotein interactions in coronavirusORGANISM assembly",
displaCy
"``ImmunogenicityNNofINaDTpolyvalentJJHIV-1NNcandidateNNvaccineNNbasedVBNonINfourteenCDwildJJtypeNNgp120NNproteinsNNSinINgoldenJJhamsters",NNS_SPpunctcasedetamodcompoundcompoundnmodcasecasenummodamodcompoundcompoundnmodprepamodpunct
displaCy
"Immunogenicity of a polyvalent HIV-1ORGANISM candidate vaccine based on fourteen wild type gp120GENE_OR_GENE_PRODUCT proteins in golden hamstersORGANISM",
displaCy
"``ADTpre-S/SNNCHO-derivedJJhepatitisNNBNNvirusNNvaccineNNprotectsVBZwoodchucksNNSfromINWHVNNproductiveJJinfection",NN_SPpunctdetcompoundamodcompoundcompoundcompoundnsubjdobjcasecompoundamodnmod
displaCy
"A pre-S/S CHO-derived hepatitis B virus vaccine protects woodchucksORGANISM from WHVORGANISM productive infection",
displaCy
"``DNANNimmunizationNNwithINtheDTcysteine-richJJinterdomainNNregionNN1CDofINtheDTPlasmodiumNNfalciparumNNvariantJJantigenNNelicitsVBZlimitedJJcross-reactiveJJantibodyNNresponses",NNS_SPpunctcompoundnsubjcasedetamodcompoundnmodnsubjcasedetcompoundcompoundamodnmodamodamodcompounddobj
displaCy
"DNACELLULAR_COMPONENT immunization with the cysteine-rich interdomain region 1 of the Plasmodium falciparum variant antigen elicits limited cross-reactive antibody responses",
displaCy
"``chimericJJlive,RBattenuatedVBNvaccine (NNChimeriVax)NNincorporatingVBGtheDTenvelopeNNgenesNNSofINJapaneseJJencephalitis (NNSA14-NN14-CD2)CDvirusNNandCCtheDTcapsidNNandCCnonstructuralJJgenesNNSofINyellowJJfever (NN17D)NNvirusNNisVBZsafe,JJimmunogenicJJandCCprotectiveJJinINnon-humanJJprimates",NNS_SPpunctamodnsubjdobjapposacldetcompounddobjcaseamodcompoundcompoundnummodagentnmodccdetconjccamodconjcaseamodcompoundcompoundnmodpredetconjconjccconjcaseamodnmod
displaCy
"chimeric live, attenuated vaccine (ChimeriVaxSIMPLE_CHEMICAL) incorporating the envelope genes of Japanese encephalitis (SA14-14-2) virusORGANISM and the capsidORGANISM and nonstructuralAMINO_ACID genes of yellow fever (17D) virus is safe, immunogenic and protective in non-human primatesORGANISM",
displaCy
"``InductionNNofINprotectiveJJimmunityNNinINanimalsNNSvaccinatedVBNwithINrecombinantJJvacciniaNNvirusesNNSthatWDTexpressVBPPreMNNandCCENNglycoproteinsNNSofINJapaneseJJencephalitisNNvirus",NN_SPpunctcaseamodnmodcasenmodaclcaseamodcompoundnmodnsubjparataxisdobjcccompoundconjcaseamodcompoundpunct
displaCy
"Induction of protective immunity in animals vaccinated with recombinant vaccinia virusesORGANISM that express PreMGENE_OR_GENE_PRODUCT and E glycoproteinsGENE_OR_GENE_PRODUCT of Japanese encephalitis virus",
displaCy
"``Long-termJJprotectionNNfromINSARSNNcoronavirusNNinfectionNNconferredVBNbyINaDTsingleJJimmunizationNNwithINanDTattenuatedVBNVSV-basedJJvaccine",NN_SPpunctamodcasecompoundcompoundnmodaclcasedetamodnmodcasedetamodamodpunct
displaCy
"Long-term protection from SARS coronavirusORGANISM infection conferred by a single immunization with an attenuated VSV-basedGENE_OR_GENE_PRODUCT vaccine",
displaCy
"``RecombinantJJmodifiedVBNvacciniaNNvirusNNAnkaraNNPexpressingVBGtheDTspikeJJglycoproteinNNofINsevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNNinducesVBZprotectiveJJneutralizingVBGantibodiesNNSprimarilyRBtargetingVBGtheDTreceptorNNbindingNNregion",NN_SPpunctamodamodcompoundcompoundnsubjacldetamoddobjcaseamodamodamodcompoundnmodamodamoddobjadvmodacldetcompoundcompoundpunct
displaCy
"Recombinant modified vaccinia virus AnkaraORGANISM expressing the spike glycoprotein of severe acute respiratory syndrome coronavirusORGANISM induces protective neutralizing antibodies primarily targeting the receptor binding region",
displaCy
"``ADTdouble-inactivatedJJwholeJJvirusNNcandidateNNSARSNNcoronavirusNNvaccineNNstimulatesVBZneutralisingVBGandCCprotectiveJJantibodyNNresponses",NNS_SPpunctdetamodamodcompoundcompoundcompoundcompoundnsubjamodccconjcompounddobj
displaCy
"A double-inactivated whole virus candidate SARS coronavirus vaccineORGANISM stimulates neutralising and protective antibody responses",
displaCy
"``InactivatedJJSARS-CoVJJvaccineNNpreparedVBNfromINwholeJJvirusNNinducesVBZaDThighJJlevelNNofINneutralizingVBGantibodiesNNSinINBALB/cNNmice",NNS_SPpunctamodamodnsubjaclcaseamodnmoddetamoddobjcaseamodnmodcasecompoundpunct
displaCy
"Inactivated SARS-CoV vaccineORGANISM prepared from whole virus induces a high level of neutralizing antibodies in BALB/c miceORGANISM",
displaCy
"``ImmunogenicityNNofINSARSNNPinactivatedVBDvaccineNNinINBALB/cNNmice",NNS_SPpunctcasecompoundamodnmodprepcompoundpunct
displaCy
"Immunogenicity of SARS inactivated vaccine in BALB/c miceORGANISM",
displaCy
"``InactivatedJJSARS-CoVJJvaccineNNelicitsVBZhighJJtitersNNSofINspikeJJprotein-specificJJantibodiesNNSthatWDTblockVBPreceptorNNbindingNNandCCvirusNNentry",NN_SPpunctamodamodnsubjamoddobjcaseamodamodnmodnsubjparataxiscompounddobjcccompoundpunct
displaCy
"Inactivated SARS-CoV vaccineORGANISM elicits high titers of spike protein-specific antibodies that block receptor binding and virus entry",
displaCy
"``SARSNNPcoronavirusNNspikeNNpolypeptideNNDNANNvaccineNNprimingVBGwithINrecombinantJJspikeNNpolypeptideNNfromINEscherichiaFWcoliFWasINboosterNNinducesVBZhighJJtiterNNofINneutralizingVBGantibodyNNagainstINSARSNNPcoronavirus",NN_SPpunctcompoundcompoundamodcompoundcompoundcompoundnsubjcaseamodamodnmodcasecompoundnmodcasenmodamoddobjcaseamodnmodcasecompoundpunct
displaCy
"SARS coronavirusORGANISM spike polypeptide DNACELLULAR_COMPONENT vaccine priming with recombinant spike polypeptide from Escherichia coliORGANISM as booster induces high titer of neutralizing antibody against SARS coronavirusORGANISM",
displaCy
"``AntigenicJJandCCimmunogenicJJcharacterizationNNofINrecombinantJJbaculovirus-expressedJJsevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNNspikeNNprotein:NNimplicationNNforINvaccineNNdesign",NN_SPpunctamodccconjcaseamodamodamodamodamodcompoundcompoundamodnmodagentcasecompoundnmod
displaCy
"Antigenic and immunogenic characterization of recombinant baculovirus-expressed severe acute respiratory syndrome coronavirusORGANISM spike protein: implication for vaccine design",
displaCy
"``EvaluationNNofINmodifiedVBNvacciniaNNvirusNNAnkaraNNPbasedVBNrecombinantJJSARSNNPvaccineNNinINferrets",NNS_SPpunctprepamodcompoundcompoundpobjamodamodcompoundpobjpreppunct
displaCy
"Evaluation of modified vaccinia virus AnkaraORGANISM based recombinant SARS vaccine in ferretsORGANISM",
displaCy
"``ExosomalJJvaccinesNNScontainingVBGtheDTSNNproteinNNofINtheDTSARSNNPcoronavirusNNinduceVBPhighJJlevelsNNSofINneutralizingVBGantibodies",NNS_SPpunctamodnsubjacldetcompounddobjcasedetcompoundnmodamoddobjcaseamodpunct
displaCy
"Exosomal vaccines containing the S protein of the SARS coronavirusORGANISM induce high levels of neutralizing antibodies",
displaCy
"``MucosalJJimmunizationNNwithINsurface-displayedJJsevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNNspikeNNproteinNNonINLactobacillusNNcaseiNNinducesVBZneutralizingVBGantibodiesNNSinINmice",NNS_SPpunctamodnsubjcaseamodamodamodamodcompoundcompoundamodnmodcasecompoundnmodamoddobjcasenmod
displaCy
"Mucosal immunization with surface-displayed severe acute respiratory syndrome coronavirusORGANISM spike protein on Lactobacillus caseiSIMPLE_CHEMICAL induces neutralizing antibodies in miceORGANISM",
displaCy
"``SwitchingNNspeciesNNStropism:NNanDTeffectiveJJwayNNtoTOmanipulateVBtheDTfelineNNcoronavirusNNgenome",NN_SPpunctcompoundcompounddetamodagentmarkacldetamodcompoundpunct
displaCy
"Switching species tropism: an effective way to manipulate the feline coronavirusORGANISM genome",
displaCy
"``AssemblyNNofINspikesNNSintoINcoronavirusNNparticlesNNSisVBZmediatedVBNbyINtheDTcarboxy-terminalJJdomainNNofINtheDTspikeNNprotein",NN_SPpunctnsubjpasspreppobjcasecompoundnmodauxpasscasedetamodnmodcasedetamodnmod
displaCy
"Assembly of spikes into coronavirus particlesORGANISM is mediated by the carboxy-terminal domain of the spike protein",
displaCy
"``Virus-likeJJparticles:NNSpassportNNtoTOimmuneJJrecognition",NN_SPpunctamodagentprepamodpunct
displaCy
"Virus-like particlesORGANISM: passport to immune recognition",
displaCy
"``HumanJJmonoclonalJJantibodyNNasINprophylaxisNNforINSARSNNcoronavirusNNinfectionNNinINferrets",NNS_SPpunctamodamodcasenmodcasecompoundcompoundnmodcasepunct
displaCy
"Human monoclonal antibodyORGANISM as prophylaxis for SARS coronavirusORGANISM infection in ferretsORGANISM",
displaCy
"``Receptor-bindingJJdomainNNofINSARS-CoVJJspikeNNproteinNNinducesVBZlong-termJJprotectiveJJimmunityNNinINanDTanimalNNmodel",NN_SPpunctamodnsubjcaseamodamodnmodamodamoddobjcasedetcompoundnmod
displaCy
"Receptor-binding domain of SARS-CoVORGANISM spike protein induces long-term protective immunity in an animal model",
displaCy
"``RecombinantJJadeno-associatedJJvirusNNexpressingVBGtheDTreceptor-bindingJJdomainNNofINsevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNNSNNproteinNNelicitsVBZneutralizingVBGantibodies:NNSimplicationNNforINdevelopingVBGSARSNNPvaccines",NNS_SPpunctamodamodnsubjacldetamoddobjcaseamodamodamodcompoundcompoundcompoundnmodamoddobjagentmarkaclcompounddobj
displaCy
"Recombinant adeno-associated virus expressing the receptor-binding domain of severe acute respiratory syndrome coronavirus SORGANISM protein elicits neutralizing antibodies: implication for developing SARS vaccines",
displaCy
"``NeutralizingVBGantibodyNNandCCprotectiveJJimmunityNNtoTOSARSNNcoronavirusNNinfectionNNofINmiceNNSinducedVBNbyINaDTsolubleJJrecombinantJJpolypeptideNNcontainingVBGanDTN-terminalJJsegmentNNofINtheDTspikeNNglycoprotein",NN_SPpunctamodccamodconjcasecompoundcompoundnmodcasenmodaclcasedetamodamodnmodacldetamoddobjcasedetamodpunct
displaCy
"Neutralizing antibody and protective immunity to SARS coronavirusORGANISM infection of miceORGANISM induced by a soluble recombinant polypeptide containing an N-terminal segment of the spike glycoprotein",
displaCy
"``TherapyNNwithINaDTsevereJJacuteJJrespiratoryJJsyndrome-associatedJJcoronavirus-neutralizingJJhumanJJmonoclonalJJantibodyNNreducesVBZdiseaseNNseverityNNandCCviralJJburdenNNinINgoldenJJSyrianJJhamsters",NNS_SPpunctnsubjcasedetamodamodamodamodamodamodamodnmodcompounddobjccamodconjprepamodamodpunct
displaCy
"Therapy with a severe acute respiratory syndrome-associated coronavirus-neutralizing human monoclonal antibodyORGANISM reduces disease severity and viral burden in golden Syrian hamstersORGANISM",
displaCy
"``VaccineNNefficacyNNinINsenescentJJmiceNNSchallengedVBNwithINrecombinantJJSARS-CoVNNbearingNNepidemicJJandCCzoonoticJJspikeNNvariants",NNS_SPpunctcompoundcaseamodnmodaclcaseamodnmodaclamodccconjamodpunct
displaCy
"VaccineORGANISM efficacy in senescent miceORGANISM challenged with recombinant SARS-CoV bearing epidemic and zoonotic spike variants",
displaCy
"``EarlyJJdeathNNafterINfelineNNinfectiousJJperitonitisNNvirusNNchallengeNNdueJJtoTOrecombinantJJvacciniaNNvirusNNimmunization",NN_SPpunctamodcaseamodamodcompoundcompoundnmodcasequantmodamodcompoundcompoundpunct
displaCy
"Early death after felineORGANISM infectious peritonitis virus challenge due to recombinant vaccinia virusORGANISM immunization",
displaCy
"``ImmunopathogenesisNNofINcoronavirusNNinfections:NNSimplicationsNNSforINSARS",NNP_SPpunctcasecompoundnmodagentcasepunct
displaCy
"Immunopathogenesis of coronavirusORGANISM infections: implications for SARS",
displaCy
"``Antibody-mediatedJJenhancementNNofINdiseaseNNinINfelineNNinfectiousJJperitonitis:NNcomparisonsNNSwithINdengueNNhemorrhagicJJfever",NN_SPpunctamodcasenmodcaseamodamodnmodagentcasecompoundamodnmod
displaCy
"Antibody-mediated enhancement of disease in felineORGANISM infectious peritonitis: comparisons with dengue hemorrhagic fever",
displaCy
"``MonoclonalJJantibodiesNNStoTOtheDTspikeJJproteinNNofINfelineNNinfectiousJJperitonitisNNvirusNNmediateVBPantibody-dependentJJenhancementNNofINinfectionNNofINfelineNNmacrophages",NNS_SPpunctamodnsubjcasedetamodnmodcaseamodamodcompoundnmodamoddobjcasenmodcaseamodpunct
displaCy
"Monoclonal antibodies to the spike protein of feline infectious peritonitis virusORGANISM mediate antibody-dependent enhancement of infection of feline macrophagesORGANISM",
displaCy
"``MonoclonalJJantibodyNNanalysisNNofINneutralizationNNandCCantibody-dependentJJenhancementNNofINfelineNNinfectiousJJperitonitisNNvirus",NN_SPpunctamodcompoundcasenmodccamodconjcaseamodamodcompoundnmod
displaCy
"Monoclonal antibody analysis of neutralization and antibody-dependent enhancement of feline infectious peritonitis virusORGANISM",
displaCy
"``EvasionNNofINantibodyNNneutralizationNNinINemergingVBGsevereJJacuteJJrespiratoryJJsyndromeNNcoronaviruses",NNS_SPpunctcasecompoundnmodcaseamodamodamodamodcompoundpunct
displaCy
"Evasion of antibody neutralization in emerging severe acute respiratory syndrome coronavirusesORGANISM",
displaCy
"``LethalJJinfectionNNofINK18-hACE2NNmiceNNSinfectedVBNwithINsevereJJacuteJJrespiratoryJJsyndromeNNcoronavirus",NN_SPpunctamodcasecompoundnmodaclcaseamodamodamodcompoundnmod
displaCy
"Lethal infection of K18-hACE2 mice infected with severe acute respiratory syndrome coronavirusORGANISM",
displaCy
"``SyntheticJJreconstructionNNofINzoonoticJJandCCearlyJJhumanJJsevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNNisolatesNNSthatWDTproduceVBPfatalJJdiseaseNNinINagedJJmice",NNS_SPpunctamodcaseamodccconjamodamodamodamodcompoundcompoundnmodnsubjparataxisamoddobjcaseamodnmod
displaCy
"Synthetic reconstruction of zoonotic and early humanORGANISM severe acute respiratory syndrome coronavirus isolatesORGANISM that produce fatal disease in aged miceORGANISM",
displaCy
"``SevereJJacuteJJrespiratoryJJsyndromeNNcoronavirusNNinfectionNNofINgoldenJJSyrianJJhamsters",NNS_SPpunctamodamodamodcompoundcompoundprepamodamodpunct
displaCy
"Severe acute respiratory syndrome coronavirusORGANISM infection of golden Syrian hamstersORGANISM",
displaCy
"``Virology:NNPSARSNNPvirusNNinfectionNNofINcatsNNSandCCferrets",NNS_SPpunctcompoundcompoundapposcasenmodccpunct
displaCy
"Virology: SARS virus infection of catsORGANISM and ferretsORGANISM",
displaCy
"``CLUSTALNNPW:NNimprovingVBGtheDTsensitivityNNofINprogressiveJJmultipleJJsequenceNNalignmentNNthroughINsequenceNNweighting,NNposition-specificJJgapNNpenaltiesNNSandCCweightNNmatrixNNchoice",NN_SPpunctcompoundagentdetdobjcaseamodamodcompoundnmodcasecompoundnmodamodcompoundconjcccompoundcompoundpunct
displaCy
"CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrixCELLULAR_COMPONENT choice",
displaCy
"``CharacterizationNNofINVLPs.NNSpunctcasenmod
displaCy
"Characterization of VLPsORGANISM.
displaCy
(-LRB-A)NNSCoVNNVLPs,NNSMHVNNVLPsNNSandCCchimericJJVLPs,NNSallDTofINwhichWDTwereVBDproducedVBNfromIN293CDTNNcells,NNSwereVBDindependentlyRBpurifiedVBNbyINsucroseNNgradientNNcentrifugationNNandCC5CDgNNofINpurifiedVBNVLPsNNSwereVBDappliedVBNtoTOeachDTlaneNNofINSDS-PAGE.NNpunctexplamodnsubjpasscompoundconjccamodconjnsubjpasscasenmodauxpassparataxiscasenummodcompoundnmodauxpassadvmodcasecompoundcompoundnmodccnummodconjcaseamodnmodauxpassconjcasedetnmodcasenmod
displaCy
(A) SCoV VLPs, MHV VLPsORGANISM and chimeric VLPsCELL, all of which were produced from 293T cellsCELL, were independently purified by sucroseSIMPLE_CHEMICAL gradient centrifugation and 5 g of purified VLPsCELL were applied to each lane of SDS-PAGE.
displaCy
ColloidalNNCoomassieNNblueJJstaining (NNCCB)NNandCCWesternNNblot (NNWB)NNanalysisNNofINpurifiedVBNSCoVNNVLPs,NNSMHVNNVLPsNNSandCCchimericJJVLPsNNSareVBPshown.VBNcompoundcompoundamodnsubjpassapposcccompoundcompoundapposconjcaseamodamodnmodcompoundconjccamodconjauxpass
displaCy
Colloidal Coomassie blue staining (CCB) and Western blot (WB) analysis of purified SCoV VLPsORGANISM, MHV VLPsORGANISM and chimeric VLPsCELL are shown.
displaCy
(-LRB-B)NNNegativeJJstainingNNofINaDTchimericJJVLP.NNpunctcompoundamodcasedetamodnmod
displaCy
(B) Negative staining of a chimeric VLPCELL.
displaCy
"``SCoV-specificJJneutralizingVBGantibodyNNtitersNNSofINimmunizedJJmiceNNSatIN28CDdaysNNSafterINimmunization (NNA)NNandCCSCoVNNtitersNNSinINlungsNNSofINmiceNNS2CDdaysNNSafterINintranasalJJchallengeNNwithIN1CD\u00d7NN10CD6CDTCIDNN50CDofINSCoVNNatINdayNN28 (CDB).NNpunctamodamodcompoundcaseamodnmodcasenummodnmodcasenmodapposccamodconjcasenmodcasenmodnummodnmodcaseamodnmodcasenummodamodnummodnummodnmodnsubjcasenmodcasenmodnsubjappos
displaCy
"SCoV-specific neutralizing antibody titers of immunized miceORGANISM at 28 days after immunization (A) and SCoVSIMPLE_CHEMICAL titers in lungsORGAN of mice 2ORGANISM days after intranasal challenge with 1 \u00d7 10 6 TCID 50 of SCoVSIMPLE_CHEMICAL at day 28 (B).
displaCy
(-LRB-A)NNMeanNNSCoV-specificJJneutralizingVBGantibodyNNtitersNNSofINtheDTmiceNNSatIN28CDdaysNNSafterINi.n.NNSpunctamodamodamodcompoundagentcasedetnmodcasenummodnmodcasenmod
displaCy
(A) Mean SCoV-specific neutralizing antibody titers of the miceORGANISM at 28 days after i.nORGANISM.
displaCy
inoculationNNofINSCoVNNorCCi.m.NNcasenmodccconj
displaCy
inoculation of SCoVSIMPLE_CHEMICAL or i.mSIMPLE_CHEMICAL.
displaCy
inoculationNNofINplacebo (NNmediumNNfromINVeroNNE6NNcells)NNSorCC2CDgNNchimericJJVLPNNsuspendedVBNinINPBSNNorCCalumNNareVBPshown.VBNnsubjpasscasenmodagentcasecompoundcompoundnmodccnummodconjamodconjaclcasenmodccconjauxpass
displaCy
inoculation of placebo (medium from Vero E6 cellsCELL) or 2 g chimeric VLPCELL suspended in PBS or alumSIMPLE_CHEMICAL are shown.
displaCy
TheDTlengthsNNSofINtheDTbarsNNSindicateVBPmeanNNvirus-specificJJserumNNneutralizingNNantibodyNNtiters.NNSdetnsubjprepdetpobjamodamodcompoundcompoundcompounddobj
displaCy
The lengths of the bars indicate mean virus-specific serumORGANISM neutralizing antibody titers.
displaCy
"''FigureNN3CDSCoV-specificJJneutralizingVBGserumNNantibodyNNtitersNNSinINtheDTimmunizedJJmiceNNSandCCSCoVNNtitersNNSinINlungsNNSofINmiceNNS2CDdaysNNSafterINintranasalJJchallengeNNwithINSCoVNNatINdayNN56.CDpunctnmodnsubjamodamodcompoundcompoundcasedetamodnmodccamodconjcasenmodcasenmodnummodnmodcaseamodnmodcasenmodcasenmodnsubj
displaCy
"Figure 3 SCoV-specific neutralizing serumORGANISM_SUBSTANCE antibody titers in the immunized miceORGANISM and SCoVSIMPLE_CHEMICAL titers in lungsORGAN of mice 2ORGANISM days after intranasal challenge with SCoVSIMPLE_CHEMICAL at day 56.
displaCy
OnINdayNN0,CDmiceNNSwereVBDeitherRBleftVBNuntreatedJJorCCwereVBDinoculatedVBNi.n.NNSpreppobjnsubjnsubjpassauxpassnmod:npmodxcompccauxpassconjdobj
displaCy
On day 0, miceORGANISM were either leftORGAN untreated or were inoculated i.nORGANISM.
displaCy
withIN1CD\u00d7NN10CD6CDTCIDNN50CDofINSCoVNNorCCinjectedJJi.m.NNcasenummodexplnummodnummodnsubjcasenmodccconjagent
displaCy
with 1 \u00d7 10 6 TCID 50 of SCoVSIMPLE_CHEMICAL or injected i.mSIMPLE_CHEMICAL.
displaCy
withINplacebo (NNmediumNNfromINVeroNNE6NNcells),NNSaDTmixtureNNofIN2CDgNNMHVNNVLPsNNSandCCPBS,NNaDTmixtureNNofIN2CDgNNchimericJJVLPNNandCCPBS,NNaDTmixtureNNofIN2CDgNNchimericJJVLPNNandCCalum,NNaDTmixtureNNofIN1CDgNNchimericJJVLPNNandCCalum,NNaDTmixtureNNofIN0.5CDgNNchimericJJVLPNNandCCalum,NNaDTmixtureNNofIN1CDgNNinfluenzaNNvirusNNvaccineNNandCCalum,NNorCCalumNNalone.RBcaseagentcasecompoundcompoundnmoddetapposcasenummodnmodcompoundagentccconjdetapposcasenummodnmodamodagentccconjdetapposcasenummodnmodamodagentccconjdetapposcasenummodnmodamodagentccconjdetapposcasenummodnmodamodagentccconjdetapposcasenummodcompoundcompoundcompoundnmodccconjccconjadvmod
displaCy
with placebo (medium from Vero E6 cellsCELL), a mixture of 2 g MHV VLPsORGANISM and PBS, a mixture of 2 g chimeric VLPCELL and PBS, a mixture of 2 g chimeric VLPCELL and alumSIMPLE_CHEMICAL, a mixture of 1 g chimeric VLPCELL and alumSIMPLE_CHEMICAL, a mixture of 0.5 g chimeric VLPCELL and alumSIMPLE_CHEMICAL, a mixture of 1 g influenza virus vaccine and alumSIMPLE_CHEMICAL, or alumSIMPLE_CHEMICAL alone.
displaCy
(-LRB-A)LSTheDTgraphNNrepresentsVBZvirus-specificJJserumNNneutralizingNNantibodyNNtitersNNSatIN28CDdaysNNSpostVBPinoculation.NNpunctexpldetnsubjamodcompoundcompoundcompounddobjcasenummodcompoundcompoundnmod
displaCy
(A) The graph represents virus-specific serumORGANISM neutralizing antibody titers at 28 days post inoculation.
displaCy
(-LRB-B)NNAtIN56CDdaysNNSbloodNNsamplesNNSwereVBDcollected,VBNandCCthenRBtheDTmice,NNSexcludingVBGthoseDTinoculatedVBNwithINliveJJSCoVNNandCCleftVBDuntreated,JJwereVBDre-inoculatedVBNwithINtheDTsameJJmaterial.NNpunctexplcasecompoundnmodcompoundnsubjpassauxpassccadvmoddetnsubjpasscasenmodaclcaseamodnmodccconjxcompauxpassconjcasedetamodnmod
displaCy
(B) At 56 days blood samplesORGANISM_SUBSTANCE were collected, and then the miceORGANISM, excluding those inoculated with live SCoVSIMPLE_CHEMICAL and leftORGAN untreated, were re-inoculated with the same material.
displaCy
TheDTgraphNNrepresentsVBZvirus-specific,JJserumneutralizingVBGantibodyNNtitersNNSatIN56CDdaysNNSafterINinitialJJinoculation.NNdetnsubjamodamodcompounddobjcasenummodnmodcaseamodnmod
displaCy
The graph represents virus-specificGENE_OR_GENE_PRODUCT, serumneutralizing antibody titers at 56 days after initial inoculation.
displaCy
(-LRB-C)NNMiceNNSwereVBDinoculatedVBNwithIN1CD\u00d7NN10CD6CDTCIDNN50CDofINSCoVNNatINdayNN56.CDpunctcompoundnsubjpassauxpasscasenummodexplnummodnummodnmodnsubjcasenmodcasenmodnsubj
displaCy
(C) Mice were inoculated with 1 \u00d7 10 6 TCID 50 of SCoVSIMPLE_CHEMICAL at day 56.
displaCy
AfterIN2CDdays,NNSmiceNNSwereVBDeuthanized,VBNandCCtheDTlungNNvirusNNtitersNNSwereVBDdetermined.VBNcasenummodnmodnsubjpassauxpassccdetcompoundcompoundnsubjpassauxpassconj
displaCy
After 2 days, miceORGANISM were euthanized, and the lung virusTISSUE titers were determined.
displaCy
TheDTverticalJJdashedVBDlineNNin (INC)NNdenotesVBZtheDTminimalJJvirusNNdetectionNNlevelNNinINthisDTassay (NN2.3CDlogNN10CDTCIDNN50CD/gNNlung).NNdetamodamodnsubjcasenmoddetamodcompoundcompounddobjcasedetnmodnummodcompoundnummodcompoundcompoundcompoundappos
displaCy
The vertical dashed line in (C) denotes the minimal virus detection level in this assay (2.3 log 10 TCID 50 /g lungORGAN).
displaCy
TheDTnumberNNofINmiceNNSusedVBNfor (INB)NNand (CCC)NNwere:VBD7CDforINplacebo,NNSCoV,NNandCC2CDgNNchimericJJVLPNNwithINPBS;NN6CDforIN2CDgNNchimericJJVLPNNwithINalum;NN5CDforINMHVNNVLPNNwithINalumNNandCC1CDgNNchimericJJVLPNNwithINalum;NN4CDforIN0.5CDgNNchimericJJVLPNNwithINalum;NNandCC3CDforINallDTotherJJgroups.NNSdetnsubjcasenmodaclcasenmodccappospredetcasenmodconjccnummodconjamodagentcasenmodagentcasenummodnmodamodnmodcasenmodagentcasecompoundnmodcasenmodccnummodconjamodnmodcasenmodnummodcasenummodnmodamodagentcasenmodccconjcasedetamodnmod
displaCy
The number of miceORGANISM used for (B) and (C) were: 7 for placebo, SCoVSIMPLE_CHEMICAL, and 2 g chimeric VLPCELL with PBS; 6 for 2 g chimeric VLPCELL with alumSIMPLE_CHEMICAL; 5 for MHV VLPORGANISM with alumSIMPLE_CHEMICAL and 1 g chimeric VLPCELL with alumSIMPLE_CHEMICAL; 4 for 0.5 g chimeric VLPCELL with alumSIMPLE_CHEMICAL; and 3 for all other groups.
displaCy
"``LungNNhistopathologyNNandCCimmunohistochemistryNNofINtheDTcontrolJJplaceboNNandCCimmunizedJJmiceNNSatIN2CDdaysNNSpostVBPSCoVNNchallenge.NNpunctcompoundnsubjccconjcasedetcompoundnmodccamodconjcasenummodcompoundamoddobj
displaCy
"LungORGAN histopathology and immunohistochemistry of the control placebo and immunized miceORGANISM at 2 days post SCoVSIMPLE_CHEMICAL challenge.
displaCy
Mice (NNSnNN=JJ5CDforINallDTgroups)NNSwereVBDinoculatedVBNwithINVeroE6NNcellNNcultureNNfluidsNNStwiceRBatINdaysNNS0CDandCC28 (CDA-C),JJimmunizedVBNwithINaDTmixtureNNofINchimericJJVLPsNNSandCCPBSNNatINdaysNNS0CDandCC28 (CDDNNandCCG),NNchimericJJVLPsNNSandCCalum (NNENNandCCH)NNatINdaysNNS0CDandCC28,CDorCCliveVBSCoVNNatINdayNN0CDonly (RBFNNandCCI).NNnsubjpassagentnummoddobjcasedetnmodauxpasscasecompoundcompoundcompoundnmodadvmodcasenmodagentccconjapposxcompcasedetnmodcaseamodnmodccconjcasenmodagentccconjagentccconjamodconjccconjagentccconjcasenmodnsubjccconjccconjxcompcasenmodnsubjnmod:npmodnmodccconj
displaCy
MiceORGANISM (n = 5 for all groups) were inoculated with VeroE6 cellCELL culture fluidsTISSUE twice at days 0 and 28 (A-C), immunized with a mixture of chimeric VLPsCELL and PBS at days 0 and 28 (D and G), chimeric VLPsCELL and alumSIMPLE_CHEMICAL (ESIMPLE_CHEMICAL and HCELL) at days 0 and 28, or live SCoVSIMPLE_CHEMICAL at day 0 only (F and IGENE_OR_GENE_PRODUCT).
displaCy
TheseDTmiceNNSwereVBDchallengedVBNwithINSCoVNNatINdayNN56CDandCCsacrificedVBNatINdayNN58.CDdetnsubjpassauxpasscasenmodcasenmodnsubjccconjcasenmodnsubj
displaCy
These miceORGANISM were challenged with SCoVSIMPLE_CHEMICAL at day 56 and sacrificed at day 58.
displaCy
(-LRB-A)NNBronchiolarJJepithelialJJcellsNNSshowedVBDswellingVBGandCCblebbingNNofINtheDTluminalJJcytoplasm,NNandCCextensiveJJcellularJJdebrisNNcomprisedVBNofINnecroticJJepitheliumNNandCCinflammatoryJJcellsNNSinINtheDTairwayNNlumen.NNpunctcompoundamodamodnsubjdobjccconjcasedetamodnmodccamodamodconjaclcaseamodnmodccamodconjcasedetcompoundnmod
displaCy
(A) Bronchiolar epithelial cellsCELL showed swelling and blebbing of the luminal cytoplasmTISSUE, and extensive cellularCELL debris comprised of necrotic epitheliumTISSUE and inflammatory cellsCELL in the airway lumenMULTI-TISSUE_STRUCTURE.
displaCy
ModerateJJperibronchiolarJJmononuclearJJinflammatoryJJcellNNinfiltratesNNSareVBPalsoRBpresent (JJhematoxylinNNandCCeosinNNstaining) (NNmagnification,NN\u00d7200).NNamodcompoundamodamodcompoundnsubjpredetadvmodcompoundccconjagentagentappos
displaCy
Moderate peribronchiolar mononuclearCELL inflammatory cell infiltrates are also present (hematoxylin and eosin staining) (magnification, \u00d7200GENE_OR_GENE_PRODUCT).
displaCy
(-LRB-B)NNThickeningNNofINtheDTbronchiolarJJinterstitialJJtissuesNNSandCCalveolarJJwallsNNSwithINmononuclearJJcellNNinfiltration (NNmagnification,NN\u00d7200).NNpunctcompoundcasedetamodamodnmodccamodconjcaseamodcompoundnmodagentappos
displaCy
(B) Thickening of the bronchiolar interstitial tissuesTISSUE and alveolar wallsTISSUE with mononuclear cellCELL infiltration (magnification, \u00d7200GENE_OR_GENE_PRODUCT).
displaCy
(-LRB-C)NNSCoVNNNNNantigenNNwasVBDdistributedVBNinINbronchiolarJJepithelialJJcells,NNSasINdeterminedVBNbyINimmunohistochemistry (NNmagnification,NN\u00d7400).NNpunctcompoundamodcompoundnsubjpassauxpasscaseamodamodnmodmarkadvclcasenmodagentappos
displaCy
(C) SCoV N antigen was distributed in bronchiolar epithelial cellsCELL, as determined by immunohistochemistry (magnification, \u00d7400GENE_OR_GENE_PRODUCT).
displaCy
(-LRB-D-F)NNBronchiolarJJepithelialJJcellsNNSshowedVBDrareJJswellingNNandCCblebbingNNofINtheDTluminalJJcytoplasm,NNandCCtheDTrareJJpresenceNNofINcellularJJdebrisNNinINairways (NNSmagnification,NN\u00d7200).NNpunctexplamodamodnsubjamoddobjccconjcasedetamodnmodccdetamodconjcaseamodnmodcasenmodagentappos
displaCy
(D-F) Bronchiolar epithelial cells showed rare swelling and blebbing of the luminal cytoplasmMULTI-TISSUE_STRUCTURE, and the rare presence of cellularCELL debris in airwaysTISSUE (magnification, \u00d7200GENE_OR_GENE_PRODUCT).
displaCy
TheDTperibronchiolarJJmononuclear,JJneutrophilNNandCCeosinophilNNinfiltrates (NNSD,NNinset) (JJmagnification,NN\u00d7400).NNdetamodagentconjcccompoundagentapposapposappos
displaCy
The peribronchiolar mononuclearCELL, neutrophilCELL and eosinophil infiltratesTISSUE (D, inset) (magnification, \u00d7400GENE_OR_GENE_PRODUCT).
displaCy
(-LRB-G-I)NNSCoVNNantigensNNSwereVBDnotRBdetectedVBNbyINimmunohistochemistryNNinINtheDTlungsNNSofINtheseDTmice (NNSmagnification,NN\u00d7400).NNpunctexplamodnsubjpassauxpassnegcasenmodcasedetnmodcasedetnmodagentappos
displaCy
(G-I) SCoV antigens were not detected by immunohistochemistry in the lungsORGAN of these miceORGANISM (magnification, \u00d7400GENE_OR_GENE_PRODUCT).
displaCy
"``AminoNNacidNNsequencesNNSofINendodomainsNNSofINSCoV,NNMHVNNandCCFIPV.NNpunctcompoundcompoundcasenmodcasenmodconjccconj
displaCy
"Amino acidAMINO_ACID sequences of endodomains of SCoVSIMPLE_CHEMICAL, MHVORGANISM and FIPVORGANISM.
displaCy
ADTCLUSTALWNNPalignmentofNNtheDTcarboxy-terminusNNofINSNNproteinNNsequencesNNSofINSCoV (NNstrainNNUrbani,NNGenBankNNaccessionNNno.NNAY278741),NNMHV (NNstrainNNA59,NNGenBankNNaccessionNNno.NNNCNN001846),CDandCCFIPV (NNGenBankNNaccessionNNno.NNAY994055).NNdetnsubjdetdobjcasecompoundcompoundnmodcasenmodcompoundagentcompoundcompoundcompoundapposconjcompoundagentcompoundcompoundcompoundapposnsubjccconjcompoundcompoundcompoundappos
displaCy
A CLUSTALW alignmentof the carboxy-terminus of S protein sequences of SCoVSIMPLE_CHEMICAL (strain Urbani, GenBank accession no. AY278741), MHVORGANISM (strain A59, GenBank accession no. NC 001846), and FIPVORGANISM (GenBank accession no. AY994055).
displaCy
TheDTtransmembraneNNdomain,NNcytoplasmicJJdomain,NNcystein-richJJregion,NNandCCcharge-richJJregionNNareVBPindicated.VBNdetamodnsubjpassamodapposamodconjccamodconjauxpass
displaCy
The transmembraneCELLULAR_COMPONENT domain, cytoplasmicORGANISM_SUBSTANCE domain, cystein-rich region, and charge-richCELLULAR_COMPONENT region are indicated.
displaCy
Charge-residuesNNSinINtheDTmembraneNNproximalJJregionNNareVBPshaded.VBNnsubjpasscasedetcompoundamodnmodauxpass
displaCy
Charge-residues in the membraneCELLULAR_COMPONENT proximal region are shaded.
displaCy
"``WePRPthankVBPDr.NNPVsevolodNNPL.NNPPopovNNPandCCotherJJmembersNNSofINtheDTElectronNNPMicroscopyNNPLaboratoryNNPandCCUniversityNNPofINTexasNNPMedicalNNPBranchNNPforINtheirPRP$excellentJJsupportNNforINelectronNNmicroscopy.NNpunctnsubjcompoundcompoundcompounddobjccamodconjprepdetcompoundcompoundpobjccconjprepcompoundcompoundpobjcasecc:preconjamodnmodcasecompoundnmod
displaCy
"We thank Dr. Vsevolod L. Popov and other members of the ElectronCELLULAR_COMPONENT Microscopy Laboratory and University of Texas Medical Branch for their excellent support for electron microscopy.
displaCy
WePRPalsoRBthankVBPSusanNNPBakerNNPforINanti-MHVNNserum,NNRobertNNPAtmarNNPforINstatisticsNNSanalysisNNandCCBobNNPCouchNNPforINvaluableJJsuggestionsNNSandCCdiscussion.NNnsubjadvmodcompounddobjprepamodpobjcompounddobjprepcompoundpobjcccompoundconjprepamodpobjccconj
displaCy
We also thank Susan Baker for anti-MHV serumORGANISM_SUBSTANCE, Robert Atmar for statistics analysis and Bob Couch for valuable suggestions and discussion.
displaCy
ThisDTworkNNwasVBDsupportedVBNbyINNationalNNPInstitutesNNPSofINHealthNNPServiceNNPContractNNPAINNP65298CDandCCgrantsNNSAI29984NNandCCAI72493.NNdetnsubjpassauxpassdativecompoundpobjprepcompoundcompoundcompoundpobjnsubjccconjapposccconj
displaCy
This work was supported by National Institutes of Health Service Contract AI 65298 and grants AI29984ORGANISM and AI72493.GENE_OR_GENE_PRODUCT
displaCy
NINNwasVBDsupportedVBNbyINaDTfellowshipNNforINlong-termJJoverseasJJresearchNNforINyoungJJinvestigatorsNNSsponsoredVBNbyINtheDTMinistryNNPofINEducation,NNPCulture,NNPSports,NNPSScienceNNPandCCTechnology,NNPJapan.NNPnsubjpassauxpassdativedetpobjprepamodamodpobjprepamodpobjacldativedetpobjpreppobjconjconjconjccconjconj
displaCy
NI was supported by a fellowship for long-term overseas research for young investigators sponsored by the Ministry of Education, Culture, Sports, Science and TechnologyGENE_OR_GENE_PRODUCT, Japan.